id,group,shortref,primrecyn,researchtype,studytype,region,country,adm1,country_return,studypop,studypop_line,studyyear,summary,aim,meth_expmatmeth_list,meth_expinfmeth_list,meth_alttotinfect_list,meth_outtotrange_list,n_total,res_expmat_symp,res_expmat_any,res_expmat_prior,res_expmat_igm,res_expmat_igg,res_expmat_prnt,res_expmat_pcrser,res_expmat_pcramn,res_expmat_trim1,res_expmat_trim2,res_expmat_trim3,res_expmat_t,res_outmat_mis,res_outmat_amn,res_expinf_symp,res_expinf_any,res_expinf_igm,res_expinf_igg,res_expinf_prnt,res_expinf_csfigm,res_expinf_pcrser,res_expinf_pcrbrain,res_expinf_pcroth,res_expinf_pcrplac,res_expinf_pcrcsf,res_expinf_ihcoth,res_outinf_micro,res_outinf_anom,res_outinf_grow,res_outinf_opht,res_outinf_oth,res_effmeas,res_pval,qual_strength,qual_weakn,qual_extvalid,qual_selectbias_cc,qual_selectbias_cs,qual_obsbias,qual_repbias,qual_confound,qual_power,causdim_temp,causdim_biol,causdim_ass,causdim_alt,causdim_cess,causdim_dose,causdim_exp,causdim_ana,causdim_spec,causdim_cons,caus_temp1,caus_temp3,caus_temp2,caus_biol1,caus_biol2,caus_biol3,caus_biol4,caus_biol5,caus_biol6,caus_biol7,caus_ass1,caus_ass2,caus_alt1,caus_alt2,caus_alt3,caus_alt4,caus_alt5,caus_alt6,caus_cess1,caus_cess2,caus_cess3,caus_dose1,caus_dose2,caus_exp1,caus_exp2,caus_exp3,caus_exp4,caus_ana1,caus_ana2,caus_ana3,caus_spec1,caus_cons1,caus_cons2,caus_cons3,caus_cons4
326,11,"Mlakar, N Engl J Med, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Brazil,Rio Grande do Norte State,Slovenia,Humans,,2015,"Woman with fever, myalgia and rash at 13 weeks gestation. Weeks 14 and 20, normal ultrasound (fetal growth and anatomy). Returned to Slovenia week 28; weeks 29 and 32, growth retardation, microcephaly and calcifications on ultrasound. Pregnancy terminated at week 32. Maternal IgG for ZIKV, DENV, YFV, WNV, TBEV positive. Autopsy, small brain stem and cerebellum, and placental calcifications. ZIKV only in fetal brain tissue by RT-PCR, flavivirus particles on electron microscopy. Fetal brain tissue negative for DENV, yellow fever virus, West Nile virus, and tick-borne encephalitis, CHIKV, lymphocytic choriomeningitis, CMV, rubella, varicella zoster virus, HSV, parvovirus B19, enteroviruses, and toxoplasmosis. Genetic syndromes excluded on clinical history. Indirect immunofluorescence revealed granular intracytoplasmic reaction in destroyed neuronal structures, which pointed to a possible location of the virus in neurons",To describe the case of an expectant mother who had a febrile illness with rash at the end of the first trimester of pregnancy while she was living in Brazil.,"symptoms/medical history, ELISA IgM (serum), ELISA IgG (serum), PRNT (serum)","ELISA IgG (serum), RT-PCR (brain tissue), RT-PCR (other tissue), IHC (brain tissue), IHC (other tissue)","CHIKV, CMV, DENV, HSV, Parvovirus B19, Rubella, TBEV, Toxoplasmosis, Varicella, WNV, YF, other","clinical microcephaly, imaging confirmed brain abnormalities, intrauterine growth restriction, ",1,1 of 1,1 of 1,0 of 1,0 of 1,1 of 1,1 of 1,,,1,,,13,,,,1 of 1,,1 of 1,,,,1 of 1,0 of 1,,,0 of 1,1 of 1,1 of 1,1 of 1,,,,,"Many congenital infections excluded; imaging of congenital brain abnormalities. Laboratory confirmed ZIKV in fetal brain, virus sequenced.",Diagnosis of maternal ZIKV infection based on self‐reported clinical symptoms. Other flavivirus infection not excluded,,,,,,,,Yes,Yes,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,,,Evidence in favour,,,,,Evidence in favour,,,,Evidence in favour,,,,,,,,,,,,,,,,Evidence in favour,,
379,15,"Sarno, PLoS Negl Trop Dis, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Brazil,Salvador,,Humans,,2016,"Pregnant woman with low fetal weight at 18 weeks gestation. Week 4, serum negative for HIV, HTLV and hepatitis C, IgM negative and IgG positive for toxoplasmosis, rubella and CMV. Week 14, ultrasound normal. No symptoms of Zika-like illness or familial congenital disorders. Weeks 26 and 30, ultrasound microcephaly, hydranencephaly and calcifications. Week 32, IUFD/Stillbirth. Autopsy showed that cerebral cortex, medulla oblongata, CSF and amniotic fluid were positive for ZIKV by RT-PCR. Heart, lung, liver, vitreous body of the eye and placenta tissue samples negative for ZIKV by RT-PCR. Time of exposure unclear. Mother did not have clinical Zika symptoms.   ","To report a case of a fetus that in addition to hydranencephaly, developed hydrops fetalis and fetal demise in association with congenital ZIKV infection. Evidence for the link between ZIKV infection and microcephaly.","symptoms/medical history, RT-PCR (amniotic fluid)","RT-PCR (brain tissue), RT-PCR (other tissue), RT-PCR (CSF)","CMV, HIV, Rubella, Toxoplasmosis, other","clinical microcephaly, imaging confirmed brain abnormalities, intrauterine growth restriction, other, ",1,0 of 1,1 of 1,0 of 1,,,,,1 of 1,,,,,,,,1 of 1,,,,,,1 of 1,0 of 1,,1 of 1,,1 of 1,1 of 1,1 of 1,,1 of 1,,,Laboratory confirmed ZIKV infection in newborn,"Time of exposure unclear (no symptoms), but assumed early T1 (normal USS week 14, first abnormal USS week 18). Mother: no laboratory diagnosis for ZIKV, Dengue, Chikungunya.  ",,,,,,,,No,Yes,No,Yes,No,No,No,No,No,Yes,,,,,,Evidence in favour,,Evidence in favour,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
395,16,"Werner, Ultrasound Obstet Gynecol, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Brazil,Rio de Janeiro,,Humans,,2015,"Clinical symptoms at 12 weeks of gestation, first and second trimester ultrasound showed normal fetal anomaly. At 37 suspection and confirmation of microcephaly. ",To report imaging findings in a 27-year-old who was referred at 12 weeks of gestation for symptoms consistent with those of ZIKV infection and fetal diagnosis of microcephaly in 37 weeks of gestation.,symptoms/medical history,,"CHIKV, Chlamydia, CMV, DENV, Rubella, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, intrauterine growth restriction, ",1,1 of 1,,,,,,,,1,,,12,,,,,,,,,,,,,,,1 of 1,1 of 1,1 of 1,,,,,"maternal TORCH Screen negative; imaging of  congenital brain abnormalities (ultrasound, MRI, CT)",Diagnosis of maternal ZIKV infection based on self‐reported clinical symptoms; no RT-PCR indentification of ZIKV; retrospective diagnosis of ZIKV infection.  ,,,,,,,,Yes,No,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
549,25,"Driggers, N Engl J Med, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Multiple countries,,United States,Humans,,2015,"A 33-year old pregnant finnish woman developed rash after stay in the Americas during gestational week 11. Serum IgM and IgG for ZIKV were positive, IgM for DENV were negative but IgG for DENV positive. YF vaccination was more than 10 years ago. US at gw13, 16 and 17 showed no evidence of microcephaly or calcifications. At gw16, ZIKV  was detected in the maternal serum. At week 20, head circumference fellt o the 24th percentile. Pregnancy was terminated at gw21. Histopathological examinations showed neuronal apoptotic foci in the neocortex but not in the basal ganglia and limbic system. The subventricular zone and white matter showed severe loss of volume and macrophages infiltration. There was no evidence of micoglial nodules commonly observed in viral encephalitis. EM analysis showed viral-like particles. No ocular anomalies were detected. The highest viral load was found in the fetal brain followed by placenta and umbilical cord, and to a lesser extent in muscle, liver, lung and spleen. Alternative infections were excluded by PCR assay. Maternal serum in gw21 was also ZIKV positive by RT-PCR, but no viral RNA was detected in the serum 11 days after termination. Viral replication was observed in cell culture (Vero E6 and SKN-SH) for virus isolated in the brain, but no virus growth in culture was observed for virus isolated in other organs. ",To present a report of a case of congenital ZIKV infection and subsequent findings in a pregnancy that was terminated at 21 weeks of gestation.,"symptoms/medical history, ELISA IgM (serum), ELISA IgG (serum), PRNT (serum), RT-PCR (serum), RT-PCR (urine), RT-PCR (amniotic fluid), RT-PCR (saliva), other","RT-PCR (brain tissue), RT-PCR (other tissue), RT-PCR (placenta/product of conception), IHC (brain tissue), other","CHIKV, CMV, DENV, HSV, Parvovirus B19, Rubella, Syphilis, Toxoplasmosis, Varicella","imaging confirmed brain abnormalities, ",1,1 of 1,1 of 1,1 of 1,1 of 1,1 of 1,1 of 1,1 of 1,1 of 1,1,,,11,,,,1 of 1,,,,,,1 of 1,1 of 1,1 of 1,,,,1 of 1,,,,,,"Diagnostic of ZIKV in mother (Serum, amniotic fluid, placenta, membrane/cord) and fetus. Isolation of ZIKV of fetal brain sample and other tissues. Infectiousness of viral particles isolated in the brain confirmed in virus cultures. ","Genetic abnormalities were not studied, or another non infectious cause.",,,,,,,,Yes,Yes,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,Evidence in favour,Evidence not in favour,Evidence in favour,Evidence in favour,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,Evidence in favour,,
615,31,"ECDC, Rapid risk assessment, 21 January 2016, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Brazil,Ceara State,,Humans,,,"Born the 18 November 2015 (residing Tejuçuoca, Ceara State). Weight: 945 grams at birth. Died within 5 min after birth. Observed lesions (US, 13 Nov 2015): microcephaly (head circumference 190 mm), fetal anasarca, polydramnios. Neonatal death five minutes after birth. Presence of Zika viral genome in blood and tissue samples of the newborn (Evandro Chagas Institute).",,,"RT-PCR (serum), RT-PCR (other tissue)",,"clinical microcephaly, abnormal amniotic fluid volume",1,,,,,,,,,,,,,,1 of 1,,1 of 1,,,,,1 of 1,,1 of 1,,,,1 of 1,,,,,,,,,,,,,,,,No,Yes,No,No,No,No,No,No,No,Yes,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,
745,36,"PAHO, Epidemiological update 31 March 2016, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Martinique,,,Humans,,2016,"First case of MC in Martinique. Diagnostics by ultrasound at 22 weeks of gestation. ZIKV found in amniotic fluid and fetal blood by PCR. Mother serology positive in blood in Jan and Feb 2016. Confirmation of microcephaly on March, 24.",,"unspecified serology, RT-PCR (amniotic fluid)",RT-PCR (serum),,"clinical microcephaly, ",1,,1 of 1,1 of 1,,,,,1 of 1,,,,,,,,,,,,,1 of 1,,,,,,1 of 1,,,,,,,,No other information,,,,,,,,Yes,Yes,No,No,No,No,No,No,No,Yes,Evidence in favour,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
922,45,"Moron, BJOG, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Brazil,,,Humans,,2016,"A pregnant 31-year old woman developed maculopapular rash without fever in the early second trimester. Fetal ultrasound had been normal at gestational week 13. Ultrasound examination at week 22 was considered normal. Ultrasound at week 29 showed signs of microcephaly (circumference > 2 SD below mean). Maternal serology was negative for DENV, syphilis, HIV, Toxoplasmosis, coxsackie B virus or indicated immunity (prior seroconversion) for rubella and CMV. RT-PCR was negative for ZIKV in the maternal serum. At week 32, the fetal head circumference was > 3 SD below the average and the brain structure abnormal (cortical atrophy, ventriculomegaly, calcifications, lissencephaly, pachygyria). The placenta was increased in thickness and showed calcifications. The baby was delivered at week 39 with an Apgar score of 9 and 10. Neonate serum was positive for Zika IgG, but negative for Zika IgM. The baby was also negative for DENV, rubella, CMV, syphilis, HIV, toxoplasmosis and hepatitis. RT-PCR of the umbilical cord was negative for ZIKV. ",To describe the clinical and radiological findings of baby born infected with Zika virus and born with microcephaly,"symptoms/medical history, RT-PCR (serum)","ELISA IgM (serum), ELISA IgG (serum), RT-PCR (other tissue)","CMV, DENV, HIV, Rubella, Syphilis, Toxoplasmosis, other","clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, abnormal amniotic fluid volume",1,1 of 1,0 of 1,0 of 0,,,,0 of 1,,,1,,,,0 of 1,,1 of 1,0 of 1,1 of 1,,,,,0 of 1,,,,1 of 1,1 of 1,,0 of 1,,,,laboratory confirmed presence of Zika antibodies,"Only IgG positivity, no evidence for Zika virus in maternal or neonate serum",,,,,,,,Yes,No,No,Yes,No,No,No,No,No,No,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
1001,51,"Culjat, Clin Infect Dis, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Brazil,,United States,Humans,,2016,"A 32-year old pregnant woman living in Pernambuco (BR) experienced maculopapular rash, conjunctivitis and arthralgia at gestational week 7. No serology was done. At gestational week 12 she moved to Hawaii (USA). At gestational week 20, fetal ultrasound revealed an asymmetric left ventricular enlargement. Follow-up ultrasound at gw 22 confirmed the ventricular anomalies. She delivered spontaneously at gw 39. Apgar scores were 7 and 8. The neonate headcircumference was 27.3 cm (< 5th percentile) indicating severe microcephaly. The infant also had overriding sutures and scalp skin rugae, as well as coronal hypospadias and bilateral cryptorchidism. The placenta showed signs of acute chorioamnionitis, deciduitis with chorangiosis and scattered intervillous thrombi in the villi. The infant experienced seizures at 5 hours of life. Ophthalmological examinations at 6 weeks of life showed cortical blindness. MRI showed less than 5 gyri in each cortex, hypoplastic corpus callosum, small brain stem, pons and medulla, intracranial calcifications, and decreased brain volume. The infant's serum tested negative for syphilis, Toxoplasma (IgG, IgM), HSV (cultures), and CMV (PCR) as well as the CSF tested negative for HSV1 and HSV2 in CSF (PCR). Infant cord blood and serum of day 7 as well as the mother's postpartum blood tested positive for ZIKV IgM by ELISA and by PRNT. Placental tissue was positive for ZIKV by RT-PCR, but negative by IHC. Maternal and infant CSF were negative for ZIKV by RT-PCR. ",To describe the first case of congenital Zika infection in the United States ,"symptoms/medical history, ELISA IgM (serum), PRNT (serum), other","ELISA IgM (serum), PRNT (serum), RT-PCR (placenta/product of conception), RT-PCR (CSF), IHC (placenta/product of conception)","CMV, HSV, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, intrauterine growth restriction, ocular disorders, ",1,1 of 1,1 of 1,0 of 1,1 of 1,,1 of 1,,,1,,,,,,,1 of 1,1 of 1,,1 of 1,,,,,1 of 1,1 of 1,,1 of 1,1 of 1,0 of 1,1 of 1,,,,"laboratory confirmed presence of Zika antibodies, and placenta positive for viral RNA",No maternal serology at time of exposure,,,,,,,,Yes,Yes,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,Evidence in favour,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,Evidence in favour,,
1075,55,"Hanaoka, bioRxiv, 2016",Yes,Clinical/epidemiological research,Case report,Americas,Brazil,"Santos, São Paulo State",,Humans,,2015,"A 32-year old pregnant woman developed febrile illness with maculopapular rash and conjunctivitis at gestational week 3-4. Two months later she tested positive for syphilis (VDRL, FTA-ABs IgG) and received treatment. She showed seroconversion (IgG positive, IgM negative) for CMV, Rubella (vaccination) and Toxoplasma gondii. Four months after the zika-related symptoms, fetal ultrasound showed minimal pericardial effusion and microcephaly. At 32 weeks of gestation, these findings were confirmed in additional ultrasounds and additional malformations such as ventriculomegaly and calficiations were detected. HIV test in the mother was negative. Karyotyping revealed a normal 46xx karyotype. Delivery at gw 38 was normal. Congenital syphilis was ruled out due to a 1:1 VDRL titer, and negative CSF and urine of the child as well as negative IHC of the placenta for syphilis antigens, DMV, adenovirus and HSV. Serology of the newborn showed IgG positivity for CMV, rubella, HSV1/2 and toxoplasma gondii. The mother also had an increase in IgG titers for toxoplasma and rubella at the time of delivery. Neuroimaging showed volumetric reduction of brain parenchyma, reduced gyration and calcifications. Saliva, urine and serum of mother and child were negative by RT-PCR for CHIKV, DENV, YF and ZIKV 3 months after birth. Maternal serum was negative for all flavivirus and CHIKV IgM (ELISA), while the newborn was IgM positive for ZIKV. The mother had neutralising antibodies to ZIKV as confirmed by PRNT and both mother and child had DENV2 PRNT. Maternal serum was PRNT negative for WNF and YF. DENV and ZIKV infection of the mother and child was confimed indirectly with a cell culture assay (C6/36). Maternal cytokine profile showed activation of IL-2 and IFN-gamma and TNF-alpha. The authors conclude that prior infection with TORCH may play a critial role in the development of Zika-related microcephaly. ",To report on a case of microcephaly diagnosed prenatally with evidence for maternal Zika infection with a background of exposure to other pathogens,"symptoms/medical history, ELISA IgM (serum), PRNT (serum), RT-PCR (serum), RT-PCR (urine), RT-PCR (saliva)","ELISA IgM (serum), ELISA IgG (serum), PRNT (serum), RT-PCR (serum), RT-PCR (urine)","CHIKV, CMV, DENV, HIV, HSV, Rubella, Syphilis, Toxoplasmosis, YF","clinical microcephaly, imaging confirmed brain abnormalities, ",1,1 of 1,1 of 1,0 of 1,0 of 1,,1 of 1,0 of 1,,1,,,,,,,1 of 1,1 of 1,1 of 1,1 of 1,,0 of 1,,,,,,1 of 1,1 of 1,,,,,,,,,,,,,,,Yes,No,No,Yes,No,No,No,No,No,No,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,Evidence in favour,,,,,,,,,,,,,,,,,,
293,10,"Schuler-Faccini, MMWR Morb Mortal Wkly Rep, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Brazil,8 states,,Humans,,2015,"35 Infants with clinical microcephaly (25 with severe microcephaly), 27 with CT or ultrasound showing brain abnormalities including calcification and 9 with lissencephaly, pachygyria or arthrogryposis. 26 mothers reported rash during pregnancy (21 in first, 5 in second trimester). Serum negative in all infants for toxoplasmosis, rubella, CMV, HSV and syphilis infections. Results of CSF testing for ZIKV pending.",To investigate the possible association of microcephaly with Zika virus infection during pregnancy and a registry for incident microcephaly cases and pregnancy outcomes among women suspected to have had Zika virus infection during pregnancy. ,symptoms/medical history,,"CMV, HSV, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, ",35,26 of 35,,,,,,,,21,5,,,,,,,,,,,,,,,,,35 of 35,27 of 27,,,,,,TORCHES excluded in infants and mothers; imaging of congenital brain abnormalities,Diagnosis of maternal ZIKV infection based on self‐reported clinical symptoms. Distinguished microcephaly (35 infants) and severe (25/35). But did not say whether severe microcephaly was more likely to be associated with abnormal CT,,,,,,,,Yes,No,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
330,12,"de Paula Freitas, JAMA Ophthalmol, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Brazil,Bahia,,Humans,,2015,"31 neonates with microcephaly and presumed ZIKV  virus congenital Infection. 2 were excluded because they did not return for assessment of alternative explanation,and 29 were included. 10/29 tested with ophthalmological abnormalities. 23/29 mothers self-reported with rash, fever or arthralgia during pregnancy. Neonates serum negative for toxoplasmosis, rubella, CMV, HSV syphilis, and HIV.","To evaluate the ocular findings in 29  infants with microcephaly associated with presumed intrauterine ZIKV infection on Salvador, Bahia, Brazil  ",symptoms/medical history,,"CMV, HIV, HSV, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, ocular disorders, ",31,23 of 29,,,,,,,,18,4,1,,,,,,,,,,,,,,,,31 of 31,,,10 of 29,,,,Several congenital infections excluded in mother,Diagnosis of maternal ZIKV infection based on self‐reported clinical symptoms. Clinical diagnosis of microcephaly    ,,,,,,,,Yes,No,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,,,,
349,13,"Martines, MMWR Morb Mortal Wkly Rep, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Brazil,Rio Grande do Norte,,Humans,,2015,"Post-mortem samples of brain tissue and placenta from 2 neonates with microcephaly, who died within 20 hours of birth. Neonatal brain tissue positive for ZIKV by RT-PCR and immunohistochemistry. Histopathology of brain included calcifications, gliosis, cell degeneration and gliosis. Other autopsy tissue samples normal. Both mothers had fever and rash during first trimester. Additional report of 2 miscarriages (11 and 13 weeks); placental tissue from 1 miscarriage showed calcification of villi.",To describes evidence of a link between Zika virus infection and microcephaly and fetal demise,symptoms/medical history,"RT-PCR (brain tissue), RT-PCR (other tissue), RT-PCR (placenta/product of conception), IHC (brain tissue), IHC (other tissue)","CMV, DENV, HIV, HSV, Rubella, Toxoplasmosis","clinical microcephaly, miscarriage",4,4 of 4,,,,,,,,4,,,,2 of 4,,,4 of 4,,,,,,2 of 2,0 of 2,2 of 3,,0 of 2,2 of 2,,,,,,,Histopathological diagnosis of congenital brain  abnormalities and testing of tissues other than brain. Testing of miscarriage tissue,Diagnosis of maternal ZIKV infection based on self-reported clinical symptoms. Other infections not investigated in mothers of neonates who died. Infections only tested for in the women with miscarriages (not reported for the women with microcephalic infants,,,,,,,,Yes,Yes,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,Evidence in favour,,Evidence in favour,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
355,14,"Calvet, Lancet Infect Dis, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Brazil,Paraiba,,Humans,,2015-2016,"Case 1, 27-year-old woman in her first pregnancy: 18 weeks gestation, itching, fever, myalgia; week 16, ultrasound normal; weeks 21 and 27 weeks, microcephaly and calcifications. Case 2, 35-year-old woman in her first pregnancy: week 10, Zika-like symptoms; week 25, microcephaly on ultrasound. Week 28, all maternal serum and urine negative for ZIKV and Zika IgM, amniotic fluid positive for ZIKV by RT-PCR and Zika IgM by ELISA. All samples negative for DENV and CHIKV by RT-PCR, and IgM/IgG negative for dengue and chikungunya, negative for toxoplasmosis, rubella, CMV, HSV, HIV, syphilis, measles, and parvovirus B19.",To detect and sequence the Zika virus genome in amniotic fluid samples of two pregnant women in Brazil whose fetuses were diagnosed with microcephaly.,"symptoms/medical history, ELISA IgM (serum), RT-PCR (serum), RT-PCR (urine), RT-PCR (amniotic fluid)",,"CHIKV, CMV, DENV, HIV, HSV, Measles, Parvovirus B19, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, ",2,2 of 2,2 of 2,0 of 2,0 of 2,,,0 of 2,2 of 2,1,1,,,,,,,,,,,,,,,,,2 of 2,2 of 2,,1 of 2,,,,"Maternal ZIKV laboratory confirmed. Many congenital infections excluded in infant; imaging of congenital brain abnormalities; laboratory confirmed ZIKV in amniotic fluid, virus sequenced.",Presumed date of infection based on self-reported clinical symptoms.,,,,,,,,Yes,Yes,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,Evidence in favour,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,,,,
271,14,"Oliveira Melo, Ultrasound Obstet Gynecol, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,Paraiba,,Humans,,2015,"Two pregnant women, fetal microcephaly, brain calcifications, CNS and ophthalmological anomalies found at 29 and 30 weeks gestation. Mothers reported symptoms, serum negative for ZIKV, amniotic fluid positive by RT-PCR. Six infants with microcephaly (with neonatal HC below the 10th percentile) diagnosed with Zika virus (test not reported), all mothers apparently symptomatic during pregnancy. Fetal ultrasound showed two cases with cerebellar involvement and three with brain calcifications. One had severe arthrogryposis.",,RT-PCR (amniotic fluid),,,"clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, ",8,,2 of 2,,,,,,2 of 2,,,,,,,,,,,,,,,,,,,8 of 8,8 of 8,,1 of 2,,,,Laboratory confirmed ZIKV in amniotic fluid; imaging of congenital brain abnormalities,Diagnosis of maternal ZIKV infection based on self‐reported clinical symptoms. Other infections not investigated,,,,,,,,Yes,Yes,No,No,No,No,No,No,No,No,Evidence in favour,,Evidence in favour,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
418,17,"Meaney-Delman, MMWR Morb Mortal Wkly Rep, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Multiple countries,,United States,Humans,,2015-2016,"Nine women who had symptoms compatible with Zika virus disease and laboratory confirmed infection during pregnancy whilst living in countries with local transmission of Zika virus. Method of exposure assessment in the mother / Range of alternative infections included detection of Zika virus, viral RNA, or viral antigen, as well as Zika IgM antibodies and Zika virus neutralizing antibody titers ≥4-fold higher than neutralizing antibody titers against dengue or other flaviviruses endemic to the region where exposure occurred. Method of exposure assessment in the fetus/neonate included RT-PCR and IHC (no other details). All followed up in the U.S. Zika virus infection during pregnancy was associated with a range of outcomes, including early pregnancy loss, microcephaly and apparently healthy infants. Six pregnant women acquired the infection during the first trimester of pregnancy, two during the second and one during the third trimester. Among the six women who reported symptoms during the first trimester there were two early pregnancy losses, two elective terminations, and delivery of a live born infant with microcephaly. Zika virus RNA was detected in both fetal remains from the women with early pregnancy loss. In one case of elective pregnancy termination, amniocentesis was performed and Zika virus RNA was detected by RT‐PCR and a fetal ultrasound previous to the termination revealed absence of corpus callosum, ventriculomegaly, and brain atrophy. In the baby born with microcephaly, Zika virus RNA was detected in the placenta by RT‐PCR and immunohistochemistry.","To summarize findings from nine women with laboratory-confirmed Zika virus infection during pregnancy who had traveled to areas with ongoing local transmission of Zika virus, including case reports for four women with various clinical outcomes.     ","symptoms/medical history, ELISA IgM (serum), PRNT (serum), RT-PCR (serum)","RT-PCR (other tissue), RT-PCR (placenta/product of conception), IHC (other tissue), IHC (placenta/product of conception)",,"clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, other, miscarriage",5,5 of 5,5 of 5,1 of 5,,,,,,5,,,,2 of 5,,,4 of 5,,,,,,,3 of 3,2 of 2,,2 of 3,1 of 5,2 of 5,,1 of 2,1 of 2,,,Some mothers acquired infection in places different from Brazil. ZIKV confirmed in mothers during pregnancy for some. ZIKV tested for in embryonic/fetal tissues    ,"Only 4/9 Mother-infant pairs reported in detail, links  to travel history are not clear. ZIKV virus infection laboratory tests performed after delivery for some mothers. No follow up of pregnancy outcomes in mothers negative for ZIKV",,,,,,,,Yes,Yes,No,No,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,
457,20,"Villamil-Gomez, Travel Med Infect Dis, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Colombia,Sucre,,Humans,,2016,"Two cases of an ongoing cohort of pregnant women with RT-PCR confirmed ZIKV infection. An abnormal outcome was detected by ultrasound only in 2 fetuses so far and consisted of brain calcifications. RT-PCR, NS1 and serology was negative for dengue and chikungunya. One of the mothers was IgG positive for toxoplasmosis, the other was bland. ",To report the preliminary findings of the first 28 pregnant women with confirmed by RT-PCR ZIKV under follow-up in the ZIKERNCOL cohort study,"symptoms/medical history, RT-PCR (serum)",,"CHIKV, CMV, DENV, HIV, HSV, Rubella, Toxoplasmosis, other","imaging confirmed brain abnormalities, ",2,2 of 2,2 of 2,2 of 2,,,,2 of 2,,,,,,,,,,,,,,,,,,,,,2 of 2,,,,,,"Laboratory confirmed ZIKV in mothers, imaging of congenital brain abnormalities; ","Missing exposure assessment in the fetus/neonate. Described as a cohort, but there will be no unexposed group, according to methods. Material of ZIKV RT-PCR not clear (Serum, Urine...); not clear if IgG and IgM tested for other infections. ",,,,,,,,Yes,No,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
542,24,"Besnard, Eurosurveillance, 2016",Yes,Clinical/epidemiological research,Case series,Pacific,French Polynesia,,,Humans,,2013-2014,"The authors reported a retrospective case series of 19 fetal and newborn cases with congenital cerebral malformations and dysfunction and detail the neurological lesions during ZIKV epidemic in French Polynesia (2013-2014). Of theses, four cases were ZIKV infection confirmed by RT-PCR in amniotic fluid. Six mother were symptomatic in the first trimester.They excluded four cases with proven aetiology (one toxo- plasmosis, two chromosomal abnormalities, one myo- pathy) and ten with usual and isolated neurological anomalies without brain damage (three intraventricular haemorrhages, three corpus callosum agenesis, one neural tube defect and three polymalformative syndromes during the first trimester of pregnancy). There was no seroconversion for CMV, hepatitis B, HIV,  rubella, syphilis and toxoplasmosis in any of the mothers; for 7 of 13 samples of amniotic fluid, PCR for EV, rubella, LCMV, HSV, VZV and DENV was negative.",To report a retrospective case series of 19 fetal and newborn cases with congenital cerebral malformations and dysfunction and detail the neurological lesions identified and the corresponding virological results.,"symptoms/medical history, RT-PCR (amniotic fluid), other",,"CMV, DENV, HIV, HSV, Rubella, Syphilis, Toxoplasmosis, Varicella, WNV, other","clinical microcephaly, imaging confirmed brain abnormalities, intrauterine growth restriction, ocular disorders, , abnormal amniotic fluid volume",19,6 of 19,4 of 7,0 of 4,,,,,4 of 7,6,,,,,3 of 19,,,,,,,,,,,,,6 of 15,18 of 19,2 of 19,1 of 1,,,,Outcome diagnostic with images. Other possible causes were discarded.,The retrospective design. Diagnostic of ZIKV infection only in seven of woman (4 were positive). The ZIKV infection was not diagnosed in fetus or newborn. Range of alternative infections were not assessed in the fetus/neonate ,,,,,,,,Yes,Yes,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,Evidence in favour,,,,,,,Evidence in favour,Evidence in favour,,,Evidence in favour,,,,,,,,,,,,,,,,,,
423,24,"Jouannic, Lancet, 2016",No,Clinical/epidemiological research,Case series,Pacific,French Polynesia,,,Humans,,2012-2016,"13 cases of microcephaly were diagnosed in 2014. Retrospectively, 6 samples of amniotic fluid were tested for ZIKV by RT-PCR of which 4 were positive. The gestational age at presumable maternal infection in these four cases was 8,9,12 weeks and unknown. ",tp report the increase of brain anomalies in Fetus/neontes due to the ZIKV epidemic between 2013 and 2014.,"symptoms/medical history, RT-PCR (amniotic fluid)",,CMV,"clinical microcephaly, imaging confirmed brain abnormalities, intrauterine growth restriction, ",13,3 of 4,4 of 6,,,,,,4 of 6,3,,,9.6700001,,,,,,,,,,,,,,,4 of 6,4 of 6,1 of 6,,,,, imaging of congenital brain abnormalities,Few other infections (only CMV) investigated; no exposure assessment in fetus/neonate; Retrospective ascertaniment and no lab confirmation of maternal infection. Not all tested for ZIKV in amniotic fluid (only available for 6/13),,,,,,,,Yes,Yes,No,Yes,No,No,No,No,No,No,Evidence in favour,,Evidence in favour,,,Evidence in favour,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
841,24,"Guillemette-Artur, Pediatr Radiol, 2016",No,Clinical/epidemiological research,Case series,Pacific,French Polynesia,,,Humans,,,"The radiological findings of three fetuses positive for Zika in amniocentesis were reviewed retrospectively. All showed microcephaly, polymicrogyria, ventriculomegaly, one also showed brainstem abnormalities",To describe and analyze the MRI findings observed in the brains of fetuses diagnosed with Zika virus infection,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Yes,No,No,Yes,No,No,No,No,No,No,Evidence in favour,,,,,,,,,,,,Evidence in favour,,,,Evidence in favour,,,,,,,,,,,,,,,,,,
568,28,"PAHO, Epidemiological update - 24 March 2016, 8 April 2016 and 28 April 2016, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Panama,,,,,,4 cases of micrcephaly reported. Case 1: report of MC and occipital encephalocele case in a newborn which died a few hours after birth. Umbilical cord positive for ZIKV by RT-PCR. Mother samples negative for ZIKV and no history of ZIKV related illness during pregnancy. Ultrasonography performed at 19 weeks showed deficit in development of neural tubes and microcephaly. Case 2: no other malformation than ʺcongenital syndrome associated with zika virusʺ. Lab confirmed ZIKV positive by rt-pct. Case 3: infant born with MC from a mother who tested positive for ZIKV in urine sample by RT-PCR. Case 4: fetus at 36 weeks of gestation diagnosed with congenital malformation by ultrasonography and lab results from mother are pending.,,"symptoms/medical history, RT-PCR (urine)",RT-PCR (other tissue),,"clinical microcephaly, imaging confirmed brain abnormalities, ",4,0 of 1,1 of 2,0 of 1,,,,,,,,,,,,,,,,,,,,1 of 1,,,,3 of 4,2 of 2,,,,,,Laboratory confirmed Zika infection,No information about temporal sequence,,,,,,,,No,Yes,No,No,No,No,No,No,No,Yes,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
718,35,"WHO, Zika situation report. Zika virus, Microcephaly and Guillain-Barré syndrome - 31 March 2016 and 07 April 2016, 2016",Yes,Clinical/epidemiological research,Case series,Africa,Cape Verde,,,Humans,,2016,"Two microcephaly cases reported. For the first case, samples from mother and infant exhibited IgG antibodies by seroneutralisation, in the second case maternal serum tested positive for Zika IgM.",,"ELISA IgM (serum), ELISA IgG (serum)",ELISA IgG (serum),,"clinical microcephaly, ",2,,2 of 2,0 of 2,1 of 1,1 of 1,,,,,,,,,,,1 of 1,,1 of 1,,,,,,,,,2 of 2,,,,,,,,No more information  ,,,,,,,,No,No,No,No,No,No,No,No,No,Yes,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,
758,37,"Microcephaly Epidemic Research Group MERG, Emerg Infect Dis, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Brazil,Pernambuco,,Humans,,2015,"In a systematic retrospective and prospective review of hospital records, 104 babies suspected to have microcephaly defined as head circumference below the third Fenton percentile for gestational age and sex were identified. Examinations included newborn serology for TORCHES (Laboratory testing was performed for 6 pathogens: serologic  testing for dengue virus; and a nontreponemal test for toxoplasmosis, rubella, cytomegalovirus,  herpes simplex virus, and syphilis) as well as imaging of the brain (CT, MRI, Ultrasound) and ophthalmological and auditory examinations as well as medical history of the mother. 70% had severe microcephaly (29 cm circumference). 2 of 25 infants with severe microcephaly had DENV IgM antibodies compared to 0 of 9 with moderate microcephaly (p=1.000). Central nervous system abnomalies included brain dysgenesis and intracranial calcifications consistent with an intrauterine infection. Radiologic abnormalities included calcifications (93%), lissencephaly, pachygyria agyria or both (96%), ventriculomegaly (60%). In a related publication (Hazin 2015, record 713), the authors reported findings of CT in 23 infants with congenital microcephaly, 7 of 7 tested infants were ZIKV IgM positive in the cerebrospinal fluid. 14 mother reported rash during pregnancy. Intracranial calcifications, ventriculomegaly, global hypogyration of the cerebral cortex were seen in all infants. Cerebellar hypoplasia was present in 17 of the  infants (74%). Three infants tested positive for syphilis, 2 for dengue, and 1 for herpes simplex virus. In another related publication (Tenorio Corderio, Lancet 2016), 30 of 31 babies born with microcephaly were positive for Zika IgM in the CSF, an 28 of 31 were Zika IgM positive in the serum. CSF and serum samples were negative for ZIKV by RT-PCR. ",To describe preliminary findings of 104 microcephaly cases in Pernambuco,symptoms/medical history,"ELISA IgM (serum), RT-PCR (serum), other","CMV, DENV, HSV, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, , ",104,59 of 100,,,,,,,,,,,,,,,30 of 31,28 of 31,,,,0 of 31,,,,,,104 of 104,104 of 104,,8 of 33,,,,Systematic sample (reviewed hospital records);  Protocol for assessment; exclusion of cases with CMV  ,"No ZIKV testing in mothers at time of clinical symptoms, no ZIKV testing in neonates; Only 7 cases with ZIKV IgM in the CSF; No data about timing of rash; No Zika surveillance data - indirect info about ZIka transmission (but see RC#460) TORCH testing for more of the case-infants; Unclear if TORCH serologies tested in mothers. Data on clinical symptoms (e.g. rash) collected post partum. ",,,,,,,,Yes,No,No,Yes,No,No,No,No,No,No,Evidence in favour,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
713,37,"Hazin, N Engl J Med, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,Pernambuco State,,Humans,,2015,"The authors reported findings of head computed tomography (CT) in 23 infants with congenital microcephaly, 7 were ZIKV IgM positive in cerebrospinal fluid. 14 mother reported rash during pregnancy. Intracranial calcifications, ventriculomegaly, global hypogyration of the cerebral cortex were seen in all infants. Cerebellar hypoplasia was present in 17 of the infants (74%).",To report findings obtained by means of head computed tomography (CT) in 23 infants with congenital microcephaly in which the clinical and epidemiologic data are compatible with congenital ZIKV infection in the Pernambuco state of Brazil.,symptoms/medical history,other,"CMV, HIV, HSV, Parvovirus B19, Rubella, Syphilis, Toxoplasmosis, Varicella","clinical microcephaly, imaging confirmed brain abnormalities, ",23,14 of 23,,,,,,,,9,5,0,,,,,,,,,,,,,,,,23 of 23,23 of 23,,,,,,Outcome evaluation with CT. TORCH infection were discarded.,"ZIKV infection diagnosed in 7 of 23 cases, by IgM in cerebrospinal fluid. The ZIKV infection in the mothers were not laboratory confirmed (only reported rash).",,,,,,,,Yes,No,No,Yes,No,No,No,No,No,No,,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
822,37,"Cavalheiro, Childs Nerv Syst, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,"Pernambuco, Maranhão, Rio Grande do Norte",,Humans,,2016,"Brain CT or MRI of 13 children with microcephaly were retrospectively reviewed. In 12 cases, the mothers had experienced Zika-related symptoms in the first trimester, in 1 case in the second trimester. All babies tested negative for toxoplasmosis, rubella, CMV, HSV and Syphilis. The main radiological findings included decreased white matter and hypoplasia of the corpus callosum, lissencephaly, increased subarachnoidal space, ventriculomegaly  and intracranial calcifications in all cases. The authors conclude that Zika infection may lead to acute destruction of the brain parenchyma and abnormal cell migration resulting in the observed pathognomic findings. In contrast to imaging findings in other viral diseases, Zika infection appears to induce gross calcifications resulting from vasculopathy and not ependymitis as observed for coxsackie infection. Lissencephaly is suggestive of exposure earlier than 18 weeks of gestation. ",To describe some radiological features in the newborns with microcephaly caused by Zika virus infection during pregnancy,symptoms/medical history,,"CMV, HSV, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, ",13,13 of 13,,,,,,,,12,1,,,,,,,,,,,,,,,,,13 of 13,13 of 13,,,,,,"imaging confirmed abnormalities, exclusion of alternative explanations","unclear overlap with MERG cases, unclear criteria for selection of cases, no laboratory confirmed Zika infection",,,,,,,,Yes,No,No,Yes,No,No,No,No,No,No,Evidence in favour,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
865,37,"Cordeiro, Lancet, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,Pernambuco,,Humans,,2015,"Serum and CSF of 31 babies born with microcephaly were analysed for Zika IgM, IgG and ZIKV, CHIKV and DENV (RT-PCR). No viral genome was detected in the samples.  Zika IgM was detected in 30 of 31 CSF samples and 28 of 31 serum samples. Monotypic response to Zika virus in the CSF was confirmed by plaque reduction  neutralisation test. Six babies had DENV IgM positivity. The authors conclude that because IgM does not cross the placenta, the antibodies must be generated by the infant as an immune response to ZIKV infection in the brain.",,,"ELISA IgM (serum), ELISA IgG (serum), IgM (CSF), RT-PCR (serum), RT-PCR (CSF)","CHIKV, DENV","clinical microcephaly, ",31,,,,,,,,,,,,,,,,30 of 31,28 of 31,,,30 of 31,,,,,,,31 of 31,,,,,,,,ZIKV IgM could also come from the maternal breast milk,,,,,,,,No,No,No,Yes,No,No,No,No,No,No,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
812,37,"de Fatima Vasco Aragao, BMJ, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,Pernambuco,,Humans,,2016,"Of 104 children diagnosed with microcephaly and/or craniofacial dysproportion at a rehabilitation center for disabled children in Pernambuco State, 23 with neuroimaging were included in a retrospective case series. The main neuroimaging findings included brain calcifications (100%), decreased brain volume (91%), malformations of cortical development (polymicrogyria, mainly frontal lobe) (95%), ventriculomegaly (86%) and cerebellum or brainstem hypoplasia (50%). ",To report radiological findings observed in computed  tomography (CT) and magnetic resonance imaging  (MRI) scans of the first cases of congenital infection  and microcephaly presumably associated with the Zika virus in the current Brazilian epidemic,symptoms/medical history,IgM (CSF),,"clinical microcephaly, imaging confirmed brain abnormalities, ",23,23 of 23,,,,,,,,17,5,0,,,,,,,,,6 of 6,,,,,,,23 of 23,23 of 23,,,,,,,,,,,,,,,Yes,No,No,No,No,No,No,No,No,No,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
439,37,"Miranda-Filho, Am J Public Health, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,,,Humans,,2016,The small proportion of cases with  laboratory confirmation reflects the difficulty  of Zika diagnosis,To provide an initial description of the congenital syndrome presumably associated with infection by Zika virus compared with other syndromes including congenital infections of established etiologies,"symptoms/medical history, RT-PCR (serum)",,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,temporality here based on symptoms,,,,,,,,Yes,No,No,No,No,No,No,No,No,No,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
914,44,"Noronha, Mem Inst Oswaldo Cruz, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Brazil,various,,Humans,,2015,"Four cases of miscarriages or newborns with microcephaly and ZIKV infections during pregnancy from northern and southern Brazil are described. The histological examinations of the placenta of one case showed hyperplasia of villous Hofbauer cells. IHC staining revealed the presence of Zika virus proteins in Hofbauer cells and some histiocytes in the intervillous space, but not in the throphoblastic epithelium. The villous samples were also positive for ZIKV by RT-PCR, but negative for DENV. The histological examination of two brain samples showed Zika virus immunopositivity by IHC in some glial cells. ",To describe histological findings in placental and brain tissues of fetuses infected with Zika at different gestational stages,symptoms/medical history,"RT-PCR (brain tissue), RT-PCR (placenta/product of conception), IHC (brain tissue), IHC (other tissue), IHC (placenta/product of conception)","CMV, HSV, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, miscarriage",4,3 of 4,,,,,,,,1,,2,,1 of 4,,,4 of 4,,,,,,1 of 1,,1 of 1,,0 of 3,3 of 4,,,,,,,,,,,,,,,,No,Yes,No,Yes,No,No,No,No,No,No,,,,,,Evidence in favour,,Evidence in favour,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,,,,
281,53,"Ventura, Lancet, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,,,Humans,,2015,"Infants with clinical microcephaly, CT confirmed cerebral calcifications and ophthalmological abnormalities: 'gross macular pigment mottling and foveal reflex loss. A well defined macular neuroretinal atrophy was detected in one child'. One mother reported rash and arthralgia in first trimester. Serum negative for toxoplasmosis, rubella, CMV, HSV, syphillis, and HIV in all mothers and infants. The headcircumference of all infants was between 28 and 28.5cm.",To report ophthalmic findings in three children with microcephaly born after the ZIKV outbreak in Brazil.,symptoms/medical history,,"CMV, HIV, HSV, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, ",3,1 of 3,,,,,,,,1,,,,,,,,,,,,,,,,,,3 of 3,3 of 3,,3 of 3,,,,"Infant TORCHES (Toxoplasmosis, rubella, CMV, herpes simplex, syphillis, and HIV) screen negative; imaging of congenital brain abnormalities",Diagnosis of maternal ZIKV infection based on self‐reported clinical symptoms. Rash in 1/3 mothers. No tests for ZIKV in neonates. ,,,,,,,,Yes,No,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
302,53,"Ventura, Arq Bras Oftalmol, 2016",No,Clinical/epidemiological research,Case series,Americas,Brazil,Pernambuco,,Humans,,,"Infants with clinical diagnosis of ZIKV vertical infection, clinical microcephaly, CT confirmed cerebral calcifications and ophthalmological findings (macular changes and optic nerve abnormalities). Seven mothers reported rash, malaise, and arthralgia during pregnancy (6 in first trimester). Serum negative in all infants for toxoplasmosis, rubella, CMV and HIV.",Assess retinal findings of infants with microcephaly and presumed ZIKV vertical infection through a complete ophthalmological exam.,symptoms/medical history,,"CMV, HIV, Rubella, Toxoplasmosis","clinical microcephaly, ocular disorders, ",10,7 of 10,,,,,,,,6,,,,,,,,,,,,,,,,,,10 of 10,,,10 of 10,,,,"Infant TORCHES (Toxoplasmosis, Rubella, CMV  and HIV) screen negative; imaging of congenital brain abnormalities",Diagnosis of maternal ZIKV infection based on self‐reported clinical symptoms. Outcome not assessed in infants (presumable intra-uterus ZIKV-infection based on negative alternative explanation assessment),,,,,,,,Yes,No,No,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
1070,54,"de Miranda, Ophthalmology, 2016",Yes,Clinical/epidemiological research,Case series,Americas,Brazil,"Pernambuco, Ceara State",,Humans,,2015,"3 children born with microcephaly underwent detailed ophthalmological examinations. The mothers had recided in Pernambuco (2) and Ceara State (1) during pregnancy and had experienced Zika-related symptoms during the first trimester. Antenatal tests in the mothers were all negative for syphilis, HIV, toxoplasma and CMV. Infant head CT demonstrated lissencephaly in all cases. Ophthalmological examinations revealed chorioretinal atrophy, pigmentary and hemorrhagic retinopathy, vascular tortuosity, absence or abnomal termination and torpedo maculopathy in all three cases. Zika testing was not performed. ",To report on 3 cases with microcephaly and expanded pigmentary and hemorrhagic retinopathy and suspected Zika virus infection,symptoms/medical history,,"CMV, HIV, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, ",3,3 of 3,,,,,,,,3,,,,,,,,,,,,,,,,,,3 of 3,3 of 3,,3 of 3,,,,"Imaging confirmed brain abnormalities, detailed ophthalmololgical assessment",No laboratory confirmed Zika infection,,,,,,,,Yes,No,No,Yes,No,No,No,No,No,No,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
419,18,"Brasil, N Engl J Med, 2016",Yes,Clinical/epidemiological research,Cohort study,Americas,Brazil,Rio de Janeiro,,Humans,,,"Cohort of 88 pregnant women with rash, 72 (88%) tested positive for ZIKV by RT‐PCR, 16 tested negative. 2 ZIKV infected women miscarried during the first trimester. 42 of ZIKV‐positive women and all ZIKV‐negative women had ultrasound scans. 12/42 (29%) ZIKV‐positive women had abnormalities vs. 0/16 (0%) of ZIKV‐negative women. Abnormalities included intrauterine growth restriction with or without microcephaly (5), cerebral calcifications (4) and abnormal amniotic fluid volume or placental blood flow (4) as well as abnormal amniotic fluid (2). Abnormalities occurred at all gestational stages. Adverse pregnancy outcomes in ZIKV infected women included 2 early pregnancy losses, 2 fetal deaths after 30 weeks, 4 live births with congenital disorders and 2 stillbirths. 88% of the women enrolled had DENV IgG. ",To describe clinical manifestations in mothers and repercussions of acute ZIKV infection in fetuses.,"symptoms/medical history, RT-PCR (serum), RT-PCR (urine)",,"CMV, DENV, Rubella, Syphilis","clinical microcephaly, imaging confirmed brain abnormalities, intrauterine growth restriction, ocular disorders, miscarriage, abnormal amniotic fluid volume",14,14 of 14,14 of 14,14 of 14,,,,,,4,6,2,20.799999,2 of 14,2 of 12,,,,,,,,,,,,,4 of 12,9 of 12,5 of 12,2 of 4,,,,Prospective cohort study supports strong association between ZIKV and adverse pregnancy outcomes. Maternal ZIKV laboratory confirmed. Complete follow up so far.,Women without ZIKV had other rash‐causing illness; other congenital infections not excluded; not all ZIKV‐infected women had ultrasound. Pregnancy outcomes for most women not yet known. No statistical measure of effect. Not clear if ZIKV -ve women had same intensity of follow up: Risk of selection bias due to differential follow-up?,Yes,,No,No,,Unclear/not reported,Unclear/not reported,Yes,No,Yes,Yes,No,No,No,No,No,Yes,Evidence in favour,,Evidence in favour,,,,,,,,Evidence in favour,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
1058,53,"Ventura, JAMA Ophthalmol, 2016",Yes,Clinical/epidemiological research,Cross-sectional study,Americas,Brazil,Pernambuco,,Humans,,2015,"Children born with microcephaly (<2 SD below average) between May and December 2015 in Recife, Pernambuco (BR) were included in a cross-sectional study conducted at the Altino Ventura Foundation in Recife. 15 cases with incomplete or positive serology for TORCH were excluded from the study, 40 cases met the inclusion criteria. All infants and their mothers underwent ophthalmological examinations. All 24 children who underwent CSF sampling tested positive for Zika IgM. 29 of 30 cases with neuroimaging showed cerebral calcifications. 67.5% of the mothers reported Zika-related symptoms during pregnancy (32.5% in the 1st trimester, 27.5% in the 2nd trimester and 7.5% in the 3rd trimester). 55% (22) of the children but no mother had ophthalmological abnormalities. Ophthalmological abnormalities included optic nerves alterations and/or macular alterations. Mothers of cases with ophthalmological alterations were more likely to have had symptoms in the first trimester than mother of cases without ophthalmological alterations. Positive serology for toxoplasmosis, rubella, syphilis, CMV, HSV and HIV was an exclusion criteria for this study. 22 of 24 children were IgM negative for DENV, two had weak positivity, but the authors concluded that the signal was due to cross-reactivity. ",,symptoms/medical history,RT-PCR (CSF),"CMV, DENV, HIV, HSV, Rubella, Syphilis, Toxoplasmosis","clinical microcephaly, imaging confirmed brain abnormalities, ocular disorders, ",40,27 of 40,,,,,,,,13,11,3,,,,,24 of 24,,,,,,,,,24 of 24,,40 of 40,29 of 30,,22 of 40,,,,,"unclear overlap with MERG study? unclear selection of cases, likely not respresentative of population",No,,,Yes,No,No,Unclear/not reported,Yes,No,No,Yes,No,No,No,No,No,No,Evidence in favour,,Evidence in favour,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,
912,43,"Paploski, Emerg Infect Dis, 2016",Yes,Clinical/epidemiological research,Ecological study/outbreak report,Americas,Brazil,Salvador,,,,,"Using data on suspected cases of Zika (as assessed by acute exanthemous illness AEI), GBS and microcephaly from the Centers for Information and Epidemiologic Surveillance of Salvador (CIES), epidemiological curves for Zika, GBS and microcephaly were reconstructed. Temporal associations between the time series were assessed by lagged time-series cross-correlations to identify the most likely time lags between Zika exposure and sequelae. The analyses showed a strong positive correlation between temporally lagged time series of Zika cases and cases with GBS or microcephaly. The time interval between Zika infection peaks and GBS peaks was 5-9 weeks. Since the GBS cases were recorded on the date of hospitalisation rather than the date when symptoms began, the authors conclude that the actual time lag between Zika infection and onset of GBS might be shorter, as observed in other outbreaks. The number of microcephaly cases peaked 30-33 weeks after the peak of AEI cases. ",To investigate the temporal associations and the time lags between the epidemiological curves and reported cases of GBS and microcephaly,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Yes,,,,,No,No,Yes,No,No,No,Yes,No,No,No,No,No,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,
531,43,"Faria, Science, 2016",No,Clinical/epidemiological research,Ecological study/outbreak report,Americas,Brazil,,,,,,"Supplementary material shows ecological analysis using surveillance data of state-level Zika cases and microcephaly cases from the Sistema de Registro de Eventos em Saúde Pública (RESP) normalised to the number of births and assessment of correlation using spearman's rank rho. The predicted most likely time point of exposure based on surveillance data was calculated as week 17 (95% CI +/-0.11 weeks) for all cases, and week 14 (95% CI +/- 0.08 weeks) for severe microcephaly cases. When location-specific Zika virus surveillance data was included in the model, the best fitting temporal lag suggested an exposure in gestational week 18 (95% CI +/- 1.58 weeks). Also, seven Brazilian Zika strains. Phylogenetic and molecular clock analysis suggested a single introduction of ZIKV in Brazil between May-December 2013. ",To quantify ZIKV evolution and explore the introduction of the virus to the Americas.,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,0.52,0.005,"Multiple methods, raw data on surveillance",Ecological analysis of surveillance data. No exclusion or assessment of alternative explanations,,,,,,,,Yes,No,Yes,No,No,No,No,No,No,No,,,Evidence in favour,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,
460,43,"Kleber de Oliveira, MMWR Morb Mortal Wkly Rep, 2016",No,Clinical/epidemiological research,Ecological study/outbreak report,Americas,Brazil,19 states,,Humans,,,"The number of expected state-level cases of microcephaly under normal circumstances were calculated from the SINASC data 2000-2014 and compared to the observed number of cases (N=574) during the epidemic using data from the Sistema de Registro de Eventos em Saúde Pública (RESP). 12 states reported microcephaly cases of 3 or more standard deviations above the historical 2000-2014 average. The states with the highest excess cases (20 SDs above average) were Bahia, Paraiba and Pernambuco. The national microcephaly prevalence in 2015 was calculated as 2 per 10,000 live births. Of note, the microcephaly prevalence in SINASC 1995-2008 was 5.1 per 10,000 births. Among 15 states with confirmed Zika infection, the prevalence rate during the epidemic was 2.8 per 10,000 live births compared to 0.6 per 10,000 live births in states without confirmed Zika transmission (p<0.001). The calculated rate ratio was 4.7, 95% CI 1.9-13.3. The prevalence in the 12 states reporting microcephaly as > 3 SDs was 4.61/10`000, corresponding to a rate ratio of 7.7 (95% CI 3.3-17.9) compared to unaffected states. The two states with the highest prevalence rates were Pernambuco (14.62; CI = 12.33-17.17) and Paraíba (10.82; CI = 8.86-13.04).   ",To examine temporal and geospatial associations between ZIka virus transmission and MC in Brazil,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,4.7 (95% CI 1.9-13.3),>0.001,Consistent and conservative case definition for MC. Include states with and without ZIKV transmission,"Ecological fallacy (limited lab evidence of ZIKV infection for the pregnancy outcomes). Data from ad hoc surveillance sytem established by the Ministry of Health after the 1st cases possibly linked to ZIKV (enhanced awarness leading to an increased ascertainment and reporting of cases, FP). MC was probably underascertained in Brazil before this event- No other possible causes of MC evaluated. If we compare the MC incidence now with MC incidence between 1995-2008, and 2000-2008 there was a decrease in MC!  ",,,,,,,,No,No,Yes,No,No,No,No,No,No,No,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,
611,43,"Reefhuis, Emerg Infect Dis, 2016",No,Clinical/epidemiological research,Ecological study/outbreak report,Americas,Brazil,Bahia,,Humans,,,"The authors generated a cohort of pregnant women using data from the live-birth information system (SINASC) and extrapolated the beginning of the pregnancy. The number of reported Zika cases was derived from published reports on acute exanthemous illness (AEI) in the city of Salvador, Bahia March-June 2015 and the epidemic curves were extrapolated for the whole Bahia State. Data on observed microcephaly cases were taken from the SINASC Sept 2015‐Feb 2016 and compared to the projections from the hypothecial cohort of pregnant women. Assuming rash was caused by Zika and pregnancy duration was 40 weeks, the data were consistent with increased exposure to ZIKV in 1st and early 2nd trimesters. ","To illustrate the expected periods of exposure and weeks of delivery for the cohorts of pregnant women potentially infected with Zika virus during outbreaks in Bahia State, Brazil.",,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Spreadsheet for public use,No individual information on pregnancy dates; no confirmed Zika cases; numbers of microcephaly cases not reported,,,,,,,,Yes,No,No,No,Yes,No,No,No,No,No,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,
994,50,"PAHO, Epidemiological update. Zika virus infection - 28 April 2016, 2016",Yes,Clinical/epidemiological research,Ecological study/outbreak report,Americas,Brazil,,,Humans,,,"up to 23Apr16: 7228 suspected cases, 3710 investigated, 1198 confirmed with evidence of congenital infection, 2320 discarded. 73%/92% suspected/confirmed cases in Northeast of Brazil. 54/251 deaths suggestive of congenital infection. plot showing increase of MC cases starting approx. 5 months after peak of first zika case epidemic in 2015 and peaking approx 9 months after the peak of the zika epidemic (personal appreciation of temporal relationship). Concurrent dengue epidemic.",,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Yes,No,No,No,Yes,No,No,No,No,No,,Evidence in favour,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,
501,21,"Cauchemez, Lancet, 2016",Yes,Clinical/epidemiological research,Modelling study,Pacific,French Polynesia,,,Humans,,,"Retrospectively constructed cohort of individual MC cases arising during the FP Epidemic in October 13-April 14, nested in ecological level population data. The analysis included a statistical model to assess the probability of exposure to Zika in a given week, based on assumed probability proportional to number of Zika cases. Tested fit of models under different assumptions about timing of period of risk. The best model yielded a presumable exposure in the first trimester. The calculated risk ratio was 53.4 (95% CI 6.5-1061.2). ",To assess the risk of microcephaly in fetuses/neonates whose mothers had been infected by Zika virus.,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"Uses multiple datasets, statistical model to combine them, sensitivity analysis for alternatives. FP data considered complete ascertainment of MC cases and good estimates of population level exposure","No individual level data linking exposed women to MC cases. Only 8 MC cases so uncertainty high.   Apparently no cases in symptomatic women. None of cases detected during outbreak (5 of 8 were terminated, so 3 cases does not give signal above expected background)    ",,,,,,,,Yes,No,Yes,No,No,No,No,No,No,No,,,Evidence in favour,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,
1049,43,"Johansson, N Engl J Med, 2016",No,Clinical/epidemiological research,Modelling study,Americas,Brazil,Bahia,,,,2015-2016,"Using data on microcephaly cases from the SINASC registry, data on suspected Zika cases from the Bahia State Secretary health Arbovirus Situation reports and baseline birth rate data from Bahia, the authors estimated the risk of microcephaly assuming Zika infection risk from 10-80% and microcephaly overreporting of 0-100%. The probability of microcephaly was calculated as the sum of the baseline risk and the risk for each trimester for a given cohort of 16,000 births. There was a strong positive assocation between the risk of microcephaly and the risk of infection in the first trimester, while the association was neglibile for the second and third trimester. The estimated risk in the first trimester ranged from 0.88% (95% CI 0.80-0.97%) with 80% Zika virus infection rate and 100% microcephaly overreporting to 13.2% (95% CI 12.0-14.4%) with 10% Zika infection rate and no overreporting of microcephaly. ",To estimate the magnitude of the risk of microcephaly in Brazil,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Yes,No,No,No,No,No,No,No,No,No,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
1,1,"Dick, Trans R Soc Trop Med Hyg, 1952",Yes,Basic and applied biomedical research,Animal experiment,Africa,Uganda,"Entebbe, Zika forest",,"Animals: Rodents, Monkeys, models: swiss mice",MR766,1947-1948,"Serum of a sentinel Rhesus monkey caught in the Zika forest was injected intracerebrally (0.03 ml) and intraperitoneally (0.06 ml) into swiss mice and one other Rhesus monkey. The intraperitoneally injected mice showed no symptoms, while the intracerebrally injected mice showed symptoms. The injected Rhesus monkey showed no symptoms. In a second experiment, virus isolates from mosquitoes caught in the zika forest were injected into a Rhesus monkey, which showed no clinical symptoms. Serum samples taken on day 8,9 and 10 were injected into mice intracerebrally, of which some mice died and one was paralyzed. The paralysis turned out to be caused by Theiler's virus. Cross-reactivity assays showed that serum taken from a Zika infected Rhesus Maquaque had no neutralising effects on the French Neurotropic strain of yellow fever, and yellow fever hyperimmune serum failed to neutralize early passage Zika virus. Cross-reactivity tests with Dengue showed that Hawaii Dengue virus immune serum had no neutralising effect on 108th mouse brain passaged Zika virus and vice versa, Zika immune serum had no significant neutralising effect on 82nd-passage Hawaii dengue virus. The same effect was also observed for Theiler's encephalomyelitis virus. The authors conclude that adult mice were relatively insusceptible to intraperitoneal inoculation of Zika virus, and that Zika is not related to yellow fever. ",To describe the isolation of yet another virus which is  believed to be hitherto unrecorded.,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"The intracerebral and subcutaneous inoculation of mice with sera of other animals is uncontrolled, other pathogens could be injected which could cause cerebral inflammation. Serial intracerebral passage is known to increase the pathogeneicity of certain viruses",,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
2,2,"Dick, Trans R Soc Trop Med Hyg, 1952",Yes,Basic and applied biomedical research,Animal experiment,Africa,Uganda,"Entebbe, Zika forest",,"Animals: Rodents, Lagomorpha, Monkeys, models: swiss mice",MR766,1947-1948,"Swiss albino mice were injected intracerebrally with virus isolates from mosquitoes caught in the Zika forest, with serum of a sentinel Rhesus monkey and with serum of Rhesus monkey inoculated with serum of the sentinel monkey. Several passages were peformed and some mice died, some were paralysed and a few recovered. After a few pasages, some mice showed evidence of motor weakness and paralysis of the limbs, usually followed by death. Occasionally, some mice with marked motor weakness recovered. An increase in passage number (=increased adaptation to mice) increased the number of paralysed mice. Infant mice of age 7 days or younger showed susceptibility to intraperitoneal injections and required a lower lethal dose (LD50), while mice 2 weeks or older were hardly infected intraperitoneally. Mice over 6 weeks of age were slightly less susceptible to intracerebral inoculation than younger mice. Adult mice could occasionally be infected intranasally with mouse-brain suspesion, but the susceptibility to this route decreased with age. Koch type experiments with visceral organs of infected mice showed that the virus did not replicate in other organs except the brain (inoculation locus), and no viraemia was observed. Cotton rats showed no symptoms of infection when injected intracerebrally with infected mouse brain suspension. Guineapigs died after intracerebral inoculation with infected mouse brain suspension, but showed no symptoms after intraperitoneal injection. Intracerebrally inoculated rabbits showed no symptoms, but their sera contained antibodies to Zika. Rhesus monkey showed no clinical symptoms (except for the sentinel monkey which had pyrexia). All monkeys inoculated with infected mouse brain suspension developed neutralising antibodies within 2 to 3 weeks after infection. The histopathology of infected mice showed signs of inflammation and neuronal degeneration in the CNS. A human seroprevalence survey (71 adults, 28 children) in Zika region, Kampala, Bwamba and West Nile region showed a total Zika positivity of 6.1%. In conclusion, the marked neurotropism in mice is in contrast with its lack in monkeys, cotton-rats, guinea pigs and rabbits.",To investigate the pathogenicity and physical properties of Zika virus. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Histopathology of mice confirms CNS lesions. ,There is little control in Koch type experiments over the inoculation with other pathogens. Serial intracerebral passage is known to increase the pathogeneicity of certain viruses,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
12,3,"Reagan, Tex Rep Biol Med, 1954",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: syrian hamster",MR766,1954,"Swiss mice were inoculated with the 146th passage of the Zika strain. After 4-8 days all mice showed neurological symptoms, mild tremor and paralysis. The brains were then harvested and suspended injected into four vaccinated and four unvaccinated mice. The unvaccinated mice showed nervous system symptoms, while the immunised mice appeared healthy. This experiment was performed to confirm the virus identity. Healthy Syrian Hamsters (18-21 days old) were inoculated intracerebrally, intranasally, intralingually, intraorally, intradermally, intrarectally, intracardially and intramuscularly with the suspesion derived from the mouse brains. Only the hamsters inoculated intracerebrally showed symptoms of the nervous system, while all other routes of inoculation did not result in symptoms. The virus was also isolated from the hamster's brain and swiss mice were inoculated with the suspension leading to nervous system symptoms in all mice. These experiments strongly suggest replication of ZIKV in syrian hamster and swiss mouse brains.",To determine the susceptibility of the Syrian Hamster to Zika and other arboviruses by various routes of exposure,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Confirmation of ZIKV as causative agent by inoculation of immunised mice; different rodent species used; different routes of exposure used,146th passage of ZIKV used: increase serial passage in mice may lead to an increased neurotropism,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,
16,4,"Bearcroft, Trans R Soc Trop Med Hyg, 1956",Yes,Basic and applied biomedical research,Animal experiment,Africa,Nigeria,,,"Animals: Rodents, models: swiss mice",Nigerian strain,1956,"A 34-year old volunteer was experimentally inoculated with Zika (Nigerian strain). The patient showed pyrexia for a few days, but experienced no sequelae and no jaundice. The virus was isolated from his serum and infant and adult swiss mice were inoculated intracerebrally and intraperitoneally, respectively. 3 of 5 adult mice showed symptoms or died after intracerebral inoculation. Histopathology reveiled encephalitis suggestive of viral infection. Serological tests were peformed for the volunteer sera. Mouse protection tests showed a rise in antibodies not only against Zika, but also against yellow fever 12 days after Zika inoculation (tested with a mouse adapted strain). HI assays showed both Zika and yellow fever antigens indicating a rise in titer on day 8 post inoculation. After the 4th week, the YF antibodies declined again. ",To explore the result of Zika virus infection in a human volunteer with particular reference to the clinical manifestations and the development of hepatitis. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Histopathological confirmation of encephalitis,Koch Type experiments are uncontrolled for other viruses,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
20,5,"Weinbren, Trans R Soc Trop Med Hyg, 1958",Yes,Basic and applied biomedical research,Animal experiment,Africa,Uganda,"Entebbe, Lunyo forest",,"Animals: Rodents, Mosquitoes, models: swiss mice?","Uganda, Lunyo forest",1956,"Two batch captures of Aedes mosquitoes was performed in Lunyno forest. Two virus strains (identified later as Zika in immunising cross-reaction assays ) were isolated and two batches of infant and adult mice were inoculated. After a few days the brains were removed and stored or used for Seitz suspension and subsequent inoculation. All of these inoculated infant mice of the first batch died and histopathological findings indicated skeletal myositis and myocarditis, and viral type encephalitis in one infant mouse. A monkey inoculated subcutaneously was shown to develop antibodies (no further information). In the second batch, one adult mouse inoculated with one Seitz filtration passage was paralysed and died. One rhesus monkey died of retroperitoneal abscess. The histopathological findings included CNS lesion such as degeneration of nerve cells, especially in the hippocampal region, nuclear swelling and pycnosis and karyorrhexis, as well as wide-spread softening of the brain and porencephaly. Other findings included skeletal muscle  (proliferation of nuclei, swollen muscle fibers, loss of striae, infiltration of phagocytes) and myocardium (necroti foci) lesions but no lesions in other organs. This effect was observed for a short-time passaged strain as well as for a long-term passaged strain. ",To report the isolation of two more strains of Zika virus from mosquitoes and to describe certain effects noted in infant mice following the inoculation of the virus. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,histopathological evidene for encephalitis in mice ,Serial intracerebral passage is known to increase the pathogeneicity of certain viruses,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
54,7,"Bell, Arch Gesamte Virusforsch, 1971",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: swiss mice",MP1751 (Uganda 1962),1971,"A lyophilized sample of ZIKV from Uganda (Entebbe, 1962) was used for intracerebral inoculation of 1-day old and five weeks old Swiss Webster mice. On day 7 all brains were harvested and suspended for further inoculation of 1-day old and 5 weeks old WS mice. After they became sick, the brains were harvested and fixed. The identification of Zika was confirmed by culture in rhesus monkey kidney cell lines and neutralization tests with Zika antisera. Histopathological findings of fixed brain slices included patchy destruction of astroglia cells, astrocyte hypertrophy both in baby mice (pyriform cells of Ammon's horn only) and 5 week old mice (whole cortex). The tissue 'presented a moth-eaten appearance'. In the cortex of infant mice, nuclei of nerve cells appeared 'empty' and nuclear chromatin particles appeared scattered. Both glial and neuronal cells showed intracytoplasmatic inclusions within networks of endoplasmatic reticulum ('viral factories'), usually closely located to the nucleus (typical appearance of group B arboviruses) and in proximity to mitochondria. Microglial cells did not appear to be affected. In the electron miscroscope examination, ZIKV was found to replicate in both astroglial cells and neurones.",To examine the histopathological changes observed after inoculation of infant and adult mice with zika virus. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Histopathologically confirmed changes.,Serial passage experiments may increase the cerebral pathogeneicity of zika. ,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
70,8,"Way, J Gen Virol, 1976",Yes,Basic and applied biomedical research,Animal experiment,Africa,Kenya,Kano Plain,,"Animals: Rodents, Cell lines: BHK-21, CE, HeLa, L cells, LLC-MK2, MA-104, MA-111, MA-134, PS-C, Vero",MR766,1975,"Several arboviruses were assessed for replication in infant (1-2 day old) and adult (3-4 weeks old) Porton strain mice and in cell culture with different cell lines. Newborn and adult mice were inoculated cerebrally with 0.01 ml 0.03 ml of virus. Samples of 0.2 ml of serial tenfold dilutions of virus were adsorbed onto cell monolayers for 1 h at 37 degrees C. To assess the neutralising activity antisera were adsorbed onto the cell culture monolayers after incubation with the virus dilutio, and also inoculated in the mice. Antisera were considered positiveif they reduced the plaque count by 80%. Suckling mice were more susceptible and yielded higher viral load for all viruses (including ZIKV) than adult mice. Zika virus showed a higher infectivity in LLC-MK2 (Rhesus Monkey Kidney cell line) than in vivo (mice). Vero cells yielded the best replication for any type of arbovirus. ZIKV replicated only in 3 cell lines (LLC-MK2, PS-C1 Pig kidney cells and Vero cells). ",To evaluate the sensitivity of in vivo and in vitro systems for measuring the neutralizing activity of antibody while at the same time comparing the effects of viruses in mice and in cell culture sytems. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Comparison of several cell lines,Secondary (culture) cells (cell lines) may express different receptors and other proteins than primary cells. ,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
502,22,"Deng, Sci China Life Sci, 2016",Yes,Basic and applied biomedical research,Animal experiment,Asia,China,,,"Animals: Rodents, models: BALB/c mice",ZIKV SZ01 (isolated from chinese traveller returning from Samoa),2016,"Two day-old BALB/c suckling mice were inoculated intracerebrally with acute phase serum of a chinese traveller returning from Samoa. Seven days post inoculation the  majority  of  the  inoculated  animals showed neurological symptoms including  inactivity,  hind-leg  paralysis  and  hypersomnia. ZIKV infection was confirmed in the brains of the mice by RT-PCR. ",,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,injection of acute phase serum of the patient instead of purified Zika virus,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
538,23,"Rossi, Am J Trop Med Hyg, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: A129, AG129, CD1 and C57B1/6J mice","FSS13025 (Asian, Cambodia)",,"Low-passage Zika virus from the Asian strain (FSS13025) was injected subcutaneously, intradermally or intraperitoneally in transgenic mice with interferon receptor IFN alpha/beta or alpha/beta/gamma deletion (A129, AG129) and in immunocompetent mice (CD1, C57B1/6J). Infected A129 and AG129 mice showed clinical signs of illness. Young mice (3 weeks) showed severe illness including tremors, and died several days post inoculation. Older animals (11 weeks) showed less severe illness and none were moribund. ZIKV was detected by Vero cell culture and IHC staining in all organs, but the highest viral loads were observed in spleen, testes and brains. Immunocompetent control mice showed no signs of illness and no viraemia was detected. Organ viral loads were not established in these models. ",To provide insight into the pathogenesis of ZIKV in contemporary animal models,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"use of mock (control, PBS) injection, use of immunocompetent and immunodeficient murine models, use of contemporary, low-passage virus strain","generalisability of immunodeficient murine models, low sample size, no assessment of viral loads in immunocompetent mice ",,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,
553,26,"Dowall, PLoS Negl Trop Dis, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: 129Sv/Ev and A129 mice",MP1751 (Uganda 1962),2016,"Mice deficient in the IFN alpha/beta receptor (A129) and wild type mice (129Sv/Ev) aged 5-6 weeks were infected subcutaneously with the african lineage of ZIKV MP1751. At day 3 and 7 post infection the animals were sacrificed and organs were harvested. Viral RNA was detected using RT-PCR and histology assays. The A129 mice showed signs of infection (reduced growth, increased temperature), but none of the wild type mice did. A129 mice showed high viral load in blood, brain, ovaries, spleen and liver both at day 3 and 6 post infection. The wild type mice had lower levels of RNA in blood, ovaries and spleen, but not in other organs. The brain of A129 mice showed inflammatory and degenerative changes in a histological preparation. These included scattered nuclear fragments, partially degenerated cells and irregularly shaped or condensed nuclei in the neurons of the hippocampus. These lesions were not observed in the wild type mice. ",To find a susceptible adult mouse model to study infection with ZIKV,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Histopathology confirms brain lesions in IFN-deficient mice,IFN k.o. mice have a natural susceptibility for viral infections and do completely not mimick the situation in humans,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,
603,30,"Dudley, bioRxiv, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,Animals: Monkeys,"H/PF/2013 (GenBank KJ776791, French Polynesia), MR766-3329 (GenBank LC002520, Uganda)",2016,"Six rhesus macaques were inoculated subcutaneously with a French Polynesian strain of Zika (3 animals) or with the African lineage (MR766) (3 animals). Viraemia was observed in all animals infected with the Asian lineage and in 2 of 3 infected with the African lineage. Compared to the African lineage, animals infected with the Asian lineage showed higher peak viraemia with the same inoculation dose. In the group exposed to the Asian lineage, ZIKV was isolated from Urine and saliva after plasma clearance. ZIKV was also detected by qRT-PCR in CSF of the Asian group suggesting viral replication in the CNS. However, infectivity of virus isolated in the CSF was not assessed. All animals showed an elevated serum creatine kinase (CK) during viraemia. ",To develop a relevant animal model to understand ZIKV pathogenesis,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"first primate model, use of a contemporary virus strain, comparison of different strains","low sample size, no direct evidence for replication of ZIKV in CNS, no evidence for structural CNS damage",,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,
626,32,"Zmurko, PLoS Negl Trop Dis, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: AG129 and SCID mice",MR766,2016,"The authors present an in vitro assay of ZIKV replication inhibition by a polymerase inhibitor (7DMA) as well as as animal model of ZIKV infection of the brain and the effect of 7DMA in vivo. 129/sv AG129 mice (k.o. for IFN receptors) were inoculated peritoneally with ZIKV (MR766). Inoculated mice showed signs of paralysis in the lower limbs. Viral RNA was detected in brain, spleen, liver and kidney as well as in testicles by RT-PCR. Histopathological analyes showed viral antigens in neurons of the brain and the spinal cord as well as in hepatocytes. The brain lesions were consistent with acute neutrophilic encephalitis. IFN-gamma and IL-18 was increased in the serum during the course of the infection. 7DMA reduced viraemia and delayed disease progression in treated animals compared to controls. SCID mice (B/T lymphocytes k.o.) showed a delayed disease progression compared to AG129 mice",To identify an inhibitor of ZIKV in vitro and to test it in a robust animal model,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Different mouse models,70 year old MR766 strain (unknown passages),,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,
779,39,"Zika experimental science team (ZEST), , 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,Animals: Monkeys,"French Polynesian strain, Asian strain",2016,"Two pregnant Indian-origin rhesus macaque (ZIKV-003 and ZIKV-005)  in the first trimester were challenged subcutaneously with 10E4 PFU French Polynesian ZIKV and 10E4 PFU Asian lineage ZIKV, respectively. Compared to male animals, the females showed prolonged viral load in the serum oscillating between 500 and 5000 copies of RNA/mL, but not in the urine. No viral RNA was detected by qRT-PCR on day 42 post infection in a sample of amniotic fluid of ZIKV-003. Initial ultrasounds of ZIKV-003 showed normal head circumference of the fetus, while later measurements at gestational day 76-81 were still in the normal range but approximately two standard deviations below average. Additionally, placental calcifications were observed. ",To assess whether fetal development is impacted by maternal infection with Asian or French Polynesian  lineage Zika virus during the first pregnancy trimester,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Prolonged viraemia points towards replication of ZIKV in the amnion or fetus. ,No results available yet on structural brain or other malformations of the fetus. Presence of ZIKV in amniotic fluid or in the fetus not verified,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,
793,40,"Lazear, Cell Host Microbe, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: Ifnar1 KO, Ifnar3 - Ifnar5 - Ifnar7 TKO, Irf3 KO, lfit1 KO, lfit2 KO, lsg15 KO, Ube1l KO, Mb21d1 KO, AG129, Tmem173 KO, Mavs KO, C57BL/6 and CD1 mice","MR766 (Uganda), Dakar41519 (Senegal), Dakar41667 (Senegal), Dakar41671 (Senegal), H/PF/2013 (French Polynesia)",2016,"Various immune-deficient mouse models as well as two wildtype (WT) models were inoculated subcutaneously or intravenously with several historic and contemporary ZIKV strains. Ifnar1 k.o. and Ifnar3/5/7 tripple k.o. mice showed signs of illness including neurological symptoms such as hindlimb weakness and paralysis, and died within 10 days post inoculation after inoculation with H/PF/2013. WT mice showed no signs of illness, or mortality. Immune-deficient mice inoculated with MR766 showed decreased mortality compared to the French Polynesian strains. Inoculation with the Dakar strains showed no mortality in adult immune-deficient or WT mice, however suckling infant WT mice died within days. Viral loads (H/PF/2013) were higher in immune-deficient Ifnar1 KO mice in spleen, liver, kidney, serum, testes, brain and spinal cord than in WT mice as detected by qRT-PCR. Viral replication was confirmed in plaque assays in testes, brain and spinal cord of Ifnar1 KO, but not WT mice. Results from other experiments were not reported. ",To assess the pathogenesis of ZIKV in various murine models with contemporary and historical strains,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"several mouse models, several ZIKV strains","No mock (control) inoculation, generalisability of transgenic mouse models, low sample size, selective reporting of results",,,,,,,,No,No,No,No,No,No,Yes,No,No,Yes,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,Evidence in favour,
973,47,"Wu, Cell Res, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: C57 mice",Isolated from a chinese patient returning from Samoa in February 2016,2016,"The ZIKV isolate was directly injected into the lateral ventricle of mouse fetuses in utero or intraperitoneally into the abdomen of pregnant mice at embryonic day 13.5. Immunostaining of infant brains harvested at day 1 showed the presence of ZIKV in the cells of the ventricular zone of the dorsal telencephalon as well as in the striatum for the directly injected model and only in the dorsal ventricular zone for the indirect model. The main cell infected were radial glial cells as shown by the colocalisation of ZIKV and BLBP proteins. ZIKV infected brains also exhibited a reduced mitotic activity suggestive of inhibition of proliferation of cortical neural progenitors. Compared to mock infected controls, ZIKV infection was associated with reduced mRNA levels of genes involved in brain development and cell cycle regulation such as MCPH1, CASC5, CKD6 and others. Contrarily, the receptor for IL17a (IL17Ra) was upregulated. Global transcriptome analysis confirmed that Zika virus infection led to the upregulation of genes involved in the immune response and cell death and to a downregulation of genes involved in cell proliferation and negative-regulation of apoptosis. Morphologically, the perimeters of the cortex of ZIKV infected mice were significantly shorter than that of controls. There were no differences in cortical thickness. ",To develop an animal model to firmly establish the link between vertical ZIKV transmission and malformation of the fetal brain,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,
985,48,"Li, Cell Stem Cell, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: ICR mice",ZIKV SZ01 (isolated from chinese traveller returning from Samoa),2016,"ZIKV SZ01 or culture medium (controls) was injected directly into the lateral ventricles of mouse embryos on day 13.5 in utero and the brains were harvested 3 to 5 days later. Breeding of pubs was unsuccessful because the mothers ate the newborns. RT-PCR detected a 300fold increase in viral RNA in the brains suggesting effective replication. Immunohistochemistry revealed ZIKV mostly in the ventricular (VZ) and subventricular zone (SVZ) where NPCs are located. Morphologically, the brains of ZIKV infected mice were smaller than that of control mice while the lateral ventricles were enlarged and VZ and SVZ were thinner. ZIKV infected brains also showed an increased activation of caspase3 mostly in the intemediate zone (IZ) and cortical plate (CP) and weakly in the SVZ as shown by IHC compared to controls, suggesting that ZIKV induces cell death mainly in immature or mature neurons. IHC also revealed to co-localisation of ZIKV and markers of progenitor cells (Sox2, Pax6, Tbr2) suggesting that ZIKV replicates mostly in NPCs. Further staining for phosphorylated H3 showed that compared to controls, ZIKV infected brains had less NPCs in the mitotic phase M, suggesting that ZIKV inhibitis the cell cycle and cell differentiation. Global transcriptome analysis (RNA-seq) showed an upregulation of genes involved in the immune response, including cytokines, and in apoptosis. Notably, AXL was also induced upon infection. On the other hand, genes involved in cell proliferation, differentiation, and cell migration were downregulated, as well as genes known to be associated with microcephaly in humans. ",To provide evidence from a mammalian model that ZIKV can cause microcephaly,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,use of controls,Direct injection of ZIKV into the lateral ventricles does not mimick natural infection nor vertical transmission,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,
986,49,"Miner, Cell, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: Infar1 +/- mice",H/FP/2013,2016,"Female Ifnar1 homozygous KO mice were crossed with wildtype males (C57BL/6) to generate Ifnar1 heterozygous offsprings. During pregnancy, the dams were inoculated subcutaneously on embryonic days 6.5 and 7.5 with the French Polynesian strain of ZIKV. The dams were sacrified 7 days later. The majority of the fetuses had died and were resorbed in utero. The remaining fetuses showed significant intrauterine growth restriction. No microcephaly was observed. High levels of viral RNA was detected by RT-PCR in the placentas as well as in the heads of the fetuses. Moreover, histological analysis showed activation of caspase3 in the mid- and hindbrains. In the dam, high levels of viral RNA was found in the brain, spleen and blood. Viral RNA was detected by FISH in placental trophoblasts, and virions were detected by electron microscopy in in the ER of mononuclear trophoblasts. Histological analyses showed destruction of placental microvasculature and apoptosis. These findings suggest that the mouse placenta is susceptible to ZIKV infection, which may lead to placental insufficiency and malfunction, and subsequent fetal growth restriction. ",To develop a mouse model for ZIKV infection and vertical transmission during pregnancy,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,,,
1029,52,"Aliota, PLoS Negl Trop Dis, 2016",Yes,Basic and applied biomedical research,Animal experiment,,,,,"Animals: Rodents, models: AG129 mice",H/PF/2013 (French Polynesia),2016,"Eight week old and three to four week old AG129 mice lacking the interferon alpha, beta and gamma receptors were inoculated subcutaneously (foot pad) or intraperitoneally with the French Polynesian ZIKV strain (H/PF/2013, log10 decreasing doses from 10E5 PFU to 1 PFU). All mice (young and adult) inoculated with 10E5 PFU ZIKV (foot pad or intraperitoneally) were all moribund and euthanized on day 7 post inoculation. Mice showed signs of lethargy, but none showed signs of paralysis during illness. Viral loads were high in all tissues (liver, spleen, brain, kidney, intestine, heart, skeletal muscle and lung), but highest in the brains of young mice as detected by qRT-PCR. Histopathological examinations showed no tissue damage in heart, liver, spleen, intestines, kidney and lung). One animal had signs of inflammation and degeneration in a hindlimb muscle sample. The brains showed major pathological changes including neutrophil infiltration of the hippocampus and adjacent to the choroid plexus, the menginges and cortical tissue, as well as necrosis and neutrophilic infiltration of primordial germ cell regions. ",To develop a murine model of ZIKV infection,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Use of a contemporary ZIKV strain,AG129 mice are not optimal models for human pathology due to the lack of IFN receptors,,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,
204,9,"Hamel, J Virol, 2015",Yes,Basic and applied biomedical research,In vitro experiment,,,,,"Cell lines: Primary human dermal fibroblasts HFF-1 and primary human epidermal keratinocytes derived from neonatal foreskins, primary peripheral blood mononuclear cells, human skin biopsy sample, HEK293T, A549, Vero cells, and A. albopictus C6/36 mosquito cells for ZIKV growth",PF-25013-18 (French Polynesia),2015,"Human primary skin fibroblasts were infected in vitro with a French Polynesia ZIKV lineage. 72 hours post infection all cells expressed ZIKV as determined by IF staining. The supernatant was used for plaque assays, and viral titers increased gradually within a few days suggestive of viral replication within the cells. The quantification of the viral RNA showed a marked increase in copy numbers within 24 hours post infection with a maximum of 10E8 RNA copies/ul in cells infected within 48 hours. A similar pattern was observed for primary human keratinocytes: Viral RNA increased markedly within 48 hours post infection with a maximum of 10^5 copies/ul. The ex vivo infection of a human skin biopsy specimen showed a gradual increase in viral RNA with a maximum at 5 days post infection suggestive of active viral replication. Histopathological findings included vacuolation and pyknotic nuclei in the stratum granulosum. HEK293T cells, transfected to express DC-SIGN, TIM-1, TIM-4, or a TAM family member (AXL or Tyro3) in a stable manner, were infected with ZIKV. The expression of either DC-SIGN, AXL or Tyro3 strongly enhanced viral infection, while the expression of TIM-1 or TIM-4 did not lead to a strong signal. A549 cells expressing TIM-1 and AXL, but not DC-SIGN were also infected with ZIKV. Viral infection was strongly inhibited when a neutralising antibody for AXL was added, while a neutralising antibody for TIM-1 did not alter infection. The same effect was observed when siRNA was applied. The abrogation of ZIKV by inhibiting or silencing AXL as also observed in the human skin biopsy specimen. The antiviral gene expression profile of infected fibroblasts showed the upregulation of TLR3, DDX58 (RIG-1), MDA5 and the CXCR3 ligand CXCL10, but no upregulation of TLR7. Inhibition of TLR3 resulted in an increased viral replication, but did not alter IFN Type 1 expression, suggesting a strong antiviral role of TLR3. Type 1 and 2 IFN was strongly upregulated in fibroblasts upon infection. Electron microscope imaging of ZIKV infected fibroblasts also showed numerous double-membrane intracytoplasmic vacuoles that are  characteristic of autophagosomes. Confocal microscopy showed the induction of the microtubule-associated protein LC3 colocalising with the expression of viral envelope proteins  The addition of an autophagy inhibitor decreased viral replication. These findings suggest ZIKV replicates via induction of autophagy in the cells.",To provide general insights into the interaction between this virus and its human host.,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"use of primary human cells and biopy specimens, parallel experiments show identical results, use of controls",in vitro results are not always applicable to in vivo situations,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,Evidence in favour,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
451,19,"Tang, Cell Stem Cell, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,Cell lines: human induced pluripotent stem cells (hiPSCs),MR766,2016,"Using an african lineage the authors tested several cell lines for permissiveness of ZIKV. The original human induced pluripotent stem cells (hiPSCs) were differentiated into forebrain-specific human neural progenitor cells (hNPCs) and neurons. Other cells under study included human embryonic stem cells (hESCs), HEK cells, human CNS cells derived from Glioblastoma and Astrocytoma, monkey IFN-deficient cells and A. Albopictus cells. The cells lines that showed 65-100% infected cells 3dpi were hNPCs, both human CNS cell lines, the monkey IFN-deficient cell line and the A. albopictus cell line. hESCs and hiPCSs showed <10% infected cells within 3 days post infection. Infectivity was tested by growing Vero cells with the supernatant of infected hNPCs, which confirmed the infectivity of ZIKV particles. ZIKV infection reduced hNPC cell viability by 29.9% and lead to a higher activation of caspase-3 compared to controls. Global transcriptome analyses showed a downregulation of genes involved in the cell-cycle and an upregulation of genes for protein transport and catabolic/apoptotic processes. These findings confirms that ZIKV infects immature neurons and suggest that ZIKV may dysregulate the cell cycle and the transcription in neural progenitor cells.",to investigate whether ZIKV directly infects human neural cells and the nature of its impact,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"Use of various CNS associated cells (progenitors, immature and mature neurons) showing consistent permissiveness","No attempt to identify receptors involved in ZIKV uptake, in vitro experiment may not be applicable to in vivo situation (induced progenitor cells may express other proteins) ",,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
647,33,"Garcez, Science, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,Cell lines: NSC derived from human iPS,MR766,2016,"hIPS cells were used to grow neural stem cells (NSC), neurospheres and brain organoids, subsequently exposed to ZIKV MR766. Compared to mock-infected cells ZIKV infected neurospheres grown from NSCs revealed presence of viral particle, had morphological abnormalities and cell detachment as well as microscopic signs of cell death such as apoptotic nuclei and swollen mitochondria. After 6 days only few neurospheres survived. ZIKV infected cerebral organoids growth area was reduced by 40% compared to mock-infected controls. After days, ZIKV infected organoids showed high induction of apoptosis as measured by caspase 3/7 activity.  These results show that ZIKV induces cell death in hIPS-derived neural stem cells and organoids. ",To study the effects of ZIKV during neural differentiation ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"Looked at different stage of human brain cells development (NSC, neurospheres, organoids)",iPS may express different proteins than cells in vivo,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
702,34,"Nowakowski, Cell Stem Cell, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,"Cell lines: HiPSC derived cells, ferret cortex cells, developing human retinal tissue",,2016,"Gene expression arrays (RNA-seq) for human developing cerebral cortex tissue showed high levels of expression for AXL and heat shock protein genes, mainly in radial glia cells, astrocytes, endothelial cells and microglia, while Tyro3 was not markedly increased. The spatial expression pattern of AXL was then determined using cerebral organoids derived from hiPSCs. Immunohistochemistry showed a strong expression in the borders of the ventricule, in radial glia-like cells and surrounding the brain capillaries of organoids at the mid-neurigenesis stage. Expression in neuronal populations were less pronounced. The examination of AXL expression at gw13.5 showed a persistent expression in radial glia and around capillaries. Expression of AXL in human retinal samples of gw10 and gw12 was found mainly in cells with a stem cell signature by mRNA seq. Immunostaining confirmed AXL expression on the outer margin of the retina. The expression of AXL was also confirmed in gliagl cells of ferret cortex. The authors propose AXL as a candidate ZIKV entry receptor in neural stem cells. 'AXL shows strong expression in human radial glia, brain capillaries and microglia. Developping human retina progenitors also show high AXL expression. AXL expression in conserved in rodents and human cerebral organoid model system'",To support the hypothesis that ZIKV use AXL receptor to enter the neural cells. To examine the expression of receptors implicated in cell entry of several enveloped virus including ZIKV across diverse cell types in the developing human brain,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Examination of the AXL expression receptors in different cell (single cells). They also looked at the expression pattern of AXL in primary human tissue sample using immunohistochemistry. They examined AXL expression from stages of early neurogenesis (GW13.5) to term.,hiPSC derived organoids may not express the same proteins as brains in vivo,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,
763,38,"Campanati, bioRxiv, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,"Cell lines: E14 swiss mouse cortex, SH-SY5Y (human neuroblastoma)","MR766, Br3189 (Brazilian)",2016,"Neurospheres were grown from different cell lines and exposed to ZIKV (African MR766 and Brazilian line). Infection with ZIKV Br led to a more pronounced shrinkage of neurospheres than infection with ZIKV MR 766. ZIKV Br infected neurospheres also exhibited a reduction in neurite processes. Differentiated infected cortical neurons showed less and simpler neurites than controls. In rodent brain cells, viral replication was more effective in the ZIKV Br lineage than in the MR766 lineage, but in neuroblastoma derived neurospheres, MR766 led to higher levels of viral RNA than ZIKV Br. Even with lower levels of ZIKV replication, Br lineage has more pronouced effecs in neurite outgrowth. These findings suggest that both lineages are neurotropic, but that ZIKV Br is more replicative in undifferentiated cells, while MR766 is more replicative in differentiated cells. Moreover, the cytokine profile of the supernatant showed that ZIKV Br upregulated IL-1beta dn TNF-alpha and certain chemokines such as RANTES, while ZIKV MR766 led to downregulation of IL-6. No infection was observed in differenciated neuronal cells with both lineages. Two modification acquired by the ZIKV Br strain may lead to differential pathogenesis: N-linked glycosylatio site at p153 of the envelope protein and mutation at cytoplasmatic loop of NS4b protein.   ",To study in vitro differences in pathogenesis between African and American (Brazil isolate) ZIKV lineage,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,Test of two ZIKV lineages and use of controls. Looked for neuropathogenesis and immune response differences.,"Use of a 70-year old virus strain from Africa, may have lost its infectious potential or may be changed due to serial passage in mouse brains",,,,,,,,No,Yes,No,No,No,No,Yes,No,No,Yes,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,Evidence in favour,
800,41,"Bayer, Cell Host Microbe, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,"Cell lines: primary human trophoblasts (PHT), trophoblast cells lines (BeWo, JEG3, HTR8), HBMEC, HT1080, U2OS, THP-1, ","MR766 (Uganda), FSS13025 (Cambodia)",2016,"Primary human trophoblasts (PHT) from full-term placentas and trophoblast cell lines were infected with the Asian and the African lineage ZIKV. After 48 hours of infection with both ZIKV strains and a DENV strain, trophoblast cell lines and HBMEC cells showed susceptibility to all viruses, but PHT cells showed no virus replication in immunofluorescence microscopy.  Moreover, levels of viral RNA were very low in PHT cells as assessed by RT-PCR. These findings suggest that ZIKV cannot infect and replicate efficiently in primary human trophoblasts. In another experiment, non-placental cells were incubated with the supernatant (conditioned medium, CM) of PHT cell cultures and infected with ZIKV. Cells exposed to CM showed a decreased infection with ZIKV and DENV compared to control cells. The same effect was observed when pre-treating the cells with IFN-beta. The induction of interferon stimulated genes (ISG) was confirmed with high-throughput microarray analyses. These and further experiments show an important antiviral role of Type III IFN (IFN lambda 1) produced by syncytiotrophoblasts and suggest that ZIKV may not infect full-term placental cells efficiently in vivo. Moreover, cells that do not express Type III IFN receptors, such as human brain microvascular endothelial cells (HBMEC) may be more susceptible to ZIKV infection. ZIKV might be transmitted not via trophoblast infection, but via infection of other placental cells such as fibroblasts or macrophages. ",To assess the ability of ZIKV to replicate in human placental trophoblasts,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"comparison of different cell lines and primary cells, different assays to show effect of type III IFNs",primary cells from full-term placentas instead of first or second trimester placentas,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,Evidence not in favour,,,,,,,,,,,,,,,,,,,,,,,,,,,,
884,42,"Qian, Cell, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,Cell lines: forebrain specific organoids grown from human iPSCs,"MR766, FSS13025 (Cambodia)",2016,"Forebrain-specific organoids were grown from human induced pluripotent stem cells (iPSCs) in a 3D spinning bioreactor. Infection with MR766 ZIKV at early organoid stage (day 14) showed decreased organoid size and reduced ventricular zone thickness suggestive of cell death and supression of neural progenitor cell (NPC) proliferation, as well as increased lumen within the ventricular structure. The infected cells were predominantly NPCs. Infection of later stage organoids (day 80) confirmed the preference for NPCs in the outer subventricular zone (oSVZ) and the ventricular zone (VZ), but also occasionally astrocytes. The infection with a contemporary asian strain from Cambodia (FSS13025) confirmed the enrichment of ZIKV in NPCs in early stage organoids. These findings suggest that ZIKV targets NPCs and causes microcephalic-like deficits in cortical development.",To model Zika exposure in the human brain in a organoid,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,3D organoid is a more realistic model of the human brain than monolayer cell cultures,"ZIKV infection assessed only with immunofluorescence (microscopically), selection process of samples for microscopy and quantification is unclear",,,,,,,,No,Yes,No,No,No,No,No,No,No,Yes,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,
947,46,"Dang, Cell Stem Cell, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,Cell lines: h9 hESC,MR766,2016,"Human embryonic stem cell (hESC) derived, immature cerebral organoids were exposed on day 10 to the African linage Zika virus (MR766). Compared to mock treated, infected organoids showed a significant decreased growth five days post infection. RT-PCR showed an increased viral copy number reflecting effective replication of Zika in the organoids. Immunostaining indicated the presence of ZIKV mainly in neural progenitor cells (NPCs). The toll-like receptor 3 (TLR3) was found upregulated in infected organoids, and treatment of organoids with a TLR3 inhibitor appeared to reserve the ZIKV-mediated shrinkage and apoptosis. These findings suggest that activation of TLR3 may play a pivotal role in the ZIKV pathogenesis of the developing brain and that ZIKV infection may activate the TLR3 pathway leading to dysregulation of neurogenesis. ",To investigate the role of ZIKV in microcephaly,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,use of organoids grown from human embryonic stem cells,use of an ancient strain (MR766) with unknown passage history and clinical relevance,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
1080,56,"Garcez, PeerJ Preprints, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,Cell lines: hiPSCs and neurospheres consisting of hiPSCS-derived neural stem cells,Brazilian strain isolated from the serum of patient in Espirito Santo (GenBank no. NC_012532),2016,"Neural stem cells induced from hiPCS were infected with the Brazilian strain ZIKV and grown to neurospheres. Compared to mock-infected controls, ZIKV infected neurospheres were significantly smaller at day 3 post inoculation (pi). ZIKV-infected neurospheres also showed reduced expression of Nestin and SOX2 suggesting a reduction in progenitor cells. Infected cells were also predominantly in a sub-G1 mitotic phase. RNA-seq analysis showed 199 downregulated and 259 upregulated proteins in ZIKV infected cells compared to controls. Downregulated proteins included proteins of the neurogenic programme and responsible for extracellular structure organisation, organelle location, synapse organisation, regulation of neuron development and cell morphogenesis. Upregulated proteins involved proteins for the cell cycle arest as well as viral replication and included in translation, cell cycle, intracellular transport, ribosome assembly. At day 6 pi, ZIKV infection led to a 50% reduction in neurosphere number, and to a complete disappearance at day 12 pi indicating cell death due to ZIKV exposure. These findings suggest that ZIKV depletes the pool of neural progenitors, reduces neuronal production through upregulation of viral translation and DNA repair as well as downregulation of the regular cell cycle and neuronal differentiation. ",To establish the transcriptome and proteome of human neurospheres infected with ZIKV,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"contemporary strain, human 3D brain models",use of induced neural cells from hiPSCs,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,,,Evidence in favour,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
1125,58,"Frumence, Virology, 2016",Yes,Basic and applied biomedical research,In vitro experiment,,,,,Cell lines: A549 (human lung epithelial cells),PF-25013-18 (French Polynesia),2016,"Human lung epithelial cells (A549) were infected with the French Polynesian strain (PF-25013-18). Successfull replication was confirmed by immunofluoresence, flow cytometry and plaque-forming assays on Vero cells. Additional incubation with an autophagy inhibitor and an autophagy inducer showed no significant change in ZIKV replication in the incubated cells. The absence of autophagy in infected cells was confirmed by western blot for marker proteins. These findings provide no evidence that the efficiency of ZIKV might be influenced by autophagy of the host cells, as suggested previously (Hamel, 2015). As shown by RT-PCR, ZIKV infection increased the transcription of interferon-stimulated genes 200fold (ISG54), cytokines such as IL-8, Il-6, Il-1beta and MCP1 and of Interferon-beta 300fold. Western blot for PARP, a marker of apoptosis, showed significant increase in apotosis in ZIKV infected cells compared to controls. Immunofluorescence showed expression of the apoptosis marker caspase-9 in ZIKV infected cells. Moreover, flow cytometry showed a high transcription of the SOD2 gene in ZIKV infected cells indicating induction of apoptosis through the accumulation of mitochondrial ROS. These findings suggest that not autophagy may not be a primary mechanism of ZIKV pathology and that ZIKV can induce apoptosis. ",To improve the knowledge on the replication of ZIKV in human cells and the host-cell responses to viral infection,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
556,27,"Homan, bioRxiv, 2016",Yes,Basic and applied biomedical research,Sequence analysis and phylogenetics,,,,,,"SPH2015/ gi 969945757 (Brazilian strain), gi 592746966 (Senegal/Cote d'Ivoire)",2016,"In silico prediction of epitopes. Polyprotein sequences of ZIKV (Brazil and African lineage) as well as of DENV, YF, WNF, TBEV, JAEV were retrieved from GenBank and curated into polyproteins, which were then segmented into individual proteins. Human and viral protein epitopes were computationally predicted. Matches were additionally filtered by the use of keywords associated with the human proteome data to select candidate pentamers for potential Zika-proteome molecular mimicry. Among the 103 pentamers identified for ZIKV, the 5 matches with the highest probability among neurological mimics were the proteins Optineurin, Synaptogyrin-1, von Willenbrand factor A, Pro-Neuropeptide Y and Neuron Navigator 2. 3D modelling of the ZIKV Brazil lineage polyprotein confirmed the matches for pro-neuropeptide Y (proNPY) (ESTEN), synaptogyrin (TESTE), neurotrophin 4 (STENS), neural cell adhesion molecule (PVITE), and VWA (TENSK) for a ZIKV region adjacent to high affinity MHC II binding site suggesting B and T-cell co-presentation. The ZIKV Brazil but not African lineage showed a mimic match for optineurin. Most lineages of DENV-3 (Brazilian lineage) were shown to mimic the mature neuropeptid Y, while the Venezuelan DENV-3 lineage has no such matches. The authors hypothesise that antibodies to ZIKV may contribute to the pathogenesis of GBS, microcephaly and ocular lesions. ",To illuminate ZIKV pathogenesis using an in silico approach and by predicting antibody mediated epitope mimicry for the human proteome. ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"3D protein prediction analysis, set of filters to detect matches, comparison of Brazilian and African lineage as well as other Viruses",,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
587,29,"Pylro, bioRxiv, 2016",Yes,Basic and applied biomedical research,Sequence analysis and phylogenetics,,,,,,,2016,"A database containing all predicted miRNAs for all ZIKV genomes on GenBank was generated. Potential human genome target sites for the miRNAs were identified. Moreover, human miRNA sequences were retrieved and mapped against the ZIKV genomes. Several rediced viral miRNAs had  targets in the human genome, which are potentially involved in brain development or function. Sequence complementarity to ZIKV was also found for human miRNAs. Several human miRNAs (hsa-miR-34a, hsa-mir-324-3p, hsa-mir-615-3p and hsa-miR-193b-3p) were identified, which target the genes CDK5/6 or WDR62. Mutations in these genes are associated with microcephaly. A complementary site for the miRNA hsa mir-21-5p, which target the gene MCHP4 associated with microcephaly, was identified in the Asian lineage (Brazil, Haiti, Martinique and French Polynesia) but not in the African lineage. ",To target mechanistic investigations of the possible relationship between ZIKV symptoms and miRNA mediated human gene expression,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,No,Yes,No,No,No,No,No,No,No,Yes,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,
1087,57,"Ganguly, bioRxiv, 2016",Yes,Basic and applied biomedical research,Sequence analysis and phylogenetics,,,,,,,,"An in silico approach using BLAST was used to find sequence similarities between the ZIKV genome sequence and human nucleotide sequences associated with microcephaly. The authors identified the gene astrotactin 2 (astn2), which shows a   high sequence alignment with the non-structural protein NS4b of ZIKV. The protein ASTN2 is expressed during developmental stages in migrating, cerebellar granule neurons and mutations of astn2 gene are associated with microcephaly in humans. The authors conclude that the ZIKV NS4b protein may alter or inhibit the physiological function of astn2 through pre-translational interactions.",To explore the parsimonous association between ZIKV and human genomes,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,No,Yes,No,No,No,No,No,No,No,No,,,,,,,,,,Evidence in favour,,,,,,,,,,,,,,,,,,,,,,,,,
47,6,"Andral, Bull World Health Organ, 1968",Yes,Environmental and vector research,Ecological/animal cross-sectional,Africa,Ethiopia,,,Animals: Monkeys,,1962-1964,"66 monkeys of the genus colobus, cynocephalus and cercopithecus were captured, killed and a serum sample was taken to assess the serological reactivity in a HI assay. 8 individuals showed evidence for Zika positivity. The brains of 23 individuals were harvested and examined. Histopathological lesions were found in 18 of 23 individuals, of which 12 had serological evidence for acute or past Zika virus infection (7 monoinfection, 2 in combination with YF and 3  in combination with group A viruses). The histopatholgical lesions included satellitosis, gliosis, involution of pyramidal cells and demyelination. All lesions indicated degenerative processes rather than acute reactive suggestive of past infections. ",To investigate the role of monkeys in the spread of yellow fever in Ethiopia,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,"Cross-reactivity of HI tests, incomplete panel for neurotropic viruses and potential for underdiagnosis of other viruses, histopathological changes not directly linked to Zika virus infection",,,,,,,,No,No,No,No,No,No,Yes,No,No,No,,,,,,,,,,,,,,,,,,,,,,,,,,Evidence in favour,,,,,,,,,