Phenotypic testing of patient herpes simplex virus type 1 and 2 isolates for acyclovir resistance by a novel method based on real-time cell analysis.

Caliaro, Oliver; Barbani, Maria Teresa; Klenja, Shkipe; Morfin, Florence; Frobert, Emilie; Gorgievski, Meri; Steinlin-Schopfer, Jacqueline; Suter-Riniker, Franziska (2020). Phenotypic testing of patient herpes simplex virus type 1 and 2 isolates for acyclovir resistance by a novel method based on real-time cell analysis. Journal of clinical virology, 125(104303), p. 104303. Elsevier 10.1016/j.jcv.2020.104303

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BACKGROUND

Acyclovir (ACV) is the most commonly used drug for herpes simplex virus (HSV) infection therapy. Prolonged antiviral therapy or prophylaxis in immunocompromised patients may promote the development of drug-resistant strains. Due to the high polymorphism in genes involved in drug resistance, phenotypic methods, although work-intensive, are still required to test drug susceptibility. Real-time cell analysis (RTCA) based methods could offer a rapid and less labor-intensive alternative for phenotypic testing of ACV resistance.

OBJECTIVE

To investigate the utility of a new RTCA based assay (RTCAA) to test acyclovir susceptibility of HSV clinical isolates.

STUDY DESIGN

Four reference strains and 93 clinical isolates (60 HSV-1 and 33 HSV-2) were tested by RTCAA. In the presence of ACV concentrations from 2.2 to 140.8 μM, Vero cells were infected with different virus dilutions. IC50 values were calculated by dose-response curve (DRC) with area-under-curve (AUC) method. The reference strains and 22 clinical isolates were additionally tested by dye-uptake assay, and IC50 values of both methods were compared.

RESULTS

IC50 values from RTCAA and dye-uptake assays were positively correlated (Spearman's rho = 0.897, p < 0.001) and quantitatively agreed (Bland-Altman plot). Based on a cut-off of 4 μM for HSV-1 and 13 μM for HSV-2, 87 isolates were classified as ACV-sensitive and 6 isolates as ACV-resistant. The reference strains showed the expected results of ACV susceptibility.

CONCLUSION

RTCAA agrees well with the dye-uptake assay. Compared with other phenotypic methods, RTCAA requires less manipulation, reduces the workload and the turnaround time, and appears to be an objective and reliable method to test ACV susceptibility.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Virology
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Infection Serology
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Molecular Biology
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Clinical Microbiology

UniBE Contributor:

Caliaro, Oliver Stephan; Barbani, Maria Teresa; Klenja, Shkipe; Gorgievski, Meri; Steinlin, Jacqueline and Suter, Franziska Marta

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1386-6532

Publisher:

Elsevier

Language:

English

Submitter:

Siegfried Hektor Hapfelmeier-Balmer

Date Deposited:

13 Jan 2021 17:45

Last Modified:

17 Jan 2021 02:54

Publisher DOI:

10.1016/j.jcv.2020.104303

PubMed ID:

32163870

Uncontrolled Keywords:

Acyclovir HSV Phenotypic assay RTCA Real-Time cell analysis Resistance

BORIS DOI:

10.48350/150236

URI:

https://boris.unibe.ch/id/eprint/150236

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