TIDE Analysis of Cryptosporidium Infections by gp60 Typing Reveals Obscured Mixed Infections
Autor: | Walter Basso, Konrad Mühlethaler, Guy Robinson, Jyothi Basapathi Raghavendra, Rachel M. Chalmers, Ines Sarah Dettwiler, Philipp Olias, Zaida Rentería-Solís, Marie-Thérèse Ruf, Sven Poppert, Mireille Meylan, Arwid Daugschies, Karin Troell, Mariko I Dale |
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Rok vydání: | 2021 |
Předmět: |
Genotype
Population Cryptosporidiosis Cryptosporidium Feces 03 medical and health sciences 0302 clinical medicine medicine Animals Humans Immunology and Allergy Parasite hosting 030212 general & internal medicine Typing education Retrospective Studies 030304 developmental biology Cryptosporidium parvum Genetics 0303 health sciences education.field_of_study biology Coinfection Transmission (medicine) Zoonosis Oocysts DNA Protozoan biology.organism_classification medicine.disease Subtyping 3. Good health Infectious Diseases Parasitic disease |
Zdroj: | The Journal of Infectious Diseases. 225:686-695 |
ISSN: | 1537-6613 0022-1899 |
Popis: | 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 Cryptosporidium parvum, 8.6% multistrain infections (13 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. Conclusions We demonstrate the value of a new, easy-to-use analytical procedure for critical characterization of C. parvum and Cryptosporidium hominis in epidemiological investigations, also applicable retrospectively. Our findings illuminate the hidden parasite diversity with important implications for tracing zoonotic and person-to-person transmissions. |
Databáze: | OpenAIRE |
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