TIDE analysis of Cryptosporidium infections by gp60 typing reveals obscured mixed infections.

Dettwiler, Ines; Troell, Karin; Robinson, Guy; Chalmers, Rachel M; Basso, Walter; Rentería-Solís, Zaida Melina; Daugschies, Arwid; Mühlethaler, Konrad; Dale, Mariko I; Basapathi, Jyothi; Ruf, Marie-Thérèse; Poppert, Sven; Meylan, Mireille; Olias, Philipp (2021). TIDE analysis of Cryptosporidium infections by gp60 typing reveals obscured mixed infections. (In Press). The journal of infectious diseases Oxford University Press 10.1093/infdis/jiab417

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BACKGROUND

Cryptosporidiosis is a parasitic disease associated with potentially fatal diarrhea. The most used method in Cryptosporidium subtyping is based on the glycoprotein gene gp60. Each infection can represent a parasite population, and it is important to investigate the influence on transmission and virulence, as well as any impact on public health investigations. However, an easy-to-use method for detection is lacking.

METHODS

Here we report on the use of the bioinformatic program TIDE for deconvolution of gp60 chromatograms. A combination of single oocyst analysis and cloning successfully confirmed the within-sample parasite population diversity. Retrospective sample analysis was conducted on archived chromatograms.

RESULTS

For C. parvum, 8.6% multi-strain infections (13 out of 152) obscured by currently used consensus base calling were detected. Importantly, we show that single oocysts can harbor a mixed population of sporozoites. We also identified a striking dominance of unappreciated polymerase stutter artefacts in all 218 chromatograms analyzed, challenging the uncritical use of gp60 typing.

DISCUSSION

We demonstrate the value of a new easy-to-use analytical procedure for critical characterization of C. parvum and C. hominis in epidemiological investigations, also applicable in retrospect. Our findings illuminate the hidden parasite diversity with important implications for tracing zoonotic and person-to-person transmissions.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases
04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Mycology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology
05 Veterinary Medicine > Department of Clinical Veterinary Medicine (DKV)
05 Veterinary Medicine > Department of Clinical Veterinary Medicine (DKV) > Clinic for Ruminants
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Animal Pathology

UniBE Contributor:

Dettwiler, Ines Sarah; Basso, Walter Ubaldo; Mühlethaler, Konrad; Dale, Mariko; Basapathi Raghavendra, Jyothi; Meylan, Mireille and Olias, Philipp Alexander

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health
500 Science > 590 Animals (Zoology)
600 Technology > 630 Agriculture

ISSN:

1537-6613

Publisher:

Oxford University Press

Language:

English

Submitter:

Nathalie Viviane Zollinger

Date Deposited:

07 Oct 2021 14:10

Last Modified:

07 Oct 2021 14:16

Publisher DOI:

10.1093/infdis/jiab417

PubMed ID:

34417806

Uncontrolled Keywords:

apicomplexa forensics protozoa stutter zoonosis

BORIS DOI:

10.48350/159766

URI:

https://boris.unibe.ch/id/eprint/159766

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