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Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers

Cyclospora cayetanensis is a human parasite transmitted via ingestion of contaminated food or water. Cases of C. cayetanensis infection acquired in the United States often go unexplained, partly because of the difficulties associated with epidemiologic investigations of such cases and the lack of ge...

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Autores principales: Hofstetter, Jessica N., Nascimento, Fernanda S., Park, Subin, Casillas, Shannon, Herwaldt, Barbara L., Arrowood, Michael J., Qvarnstrom, Yvonne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: EDP Sciences 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6354607/
https://www.ncbi.nlm.nih.gov/pubmed/30702060
http://dx.doi.org/10.1051/parasite/2019004
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author Hofstetter, Jessica N.
Nascimento, Fernanda S.
Park, Subin
Casillas, Shannon
Herwaldt, Barbara L.
Arrowood, Michael J.
Qvarnstrom, Yvonne
author_facet Hofstetter, Jessica N.
Nascimento, Fernanda S.
Park, Subin
Casillas, Shannon
Herwaldt, Barbara L.
Arrowood, Michael J.
Qvarnstrom, Yvonne
author_sort Hofstetter, Jessica N.
collection PubMed
description Cyclospora cayetanensis is a human parasite transmitted via ingestion of contaminated food or water. Cases of C. cayetanensis infection acquired in the United States often go unexplained, partly because of the difficulties associated with epidemiologic investigations of such cases and the lack of genotyping methods. A Multilocus Sequence Typing (MLST) method for C. cayetanensis based on five microsatellite loci amplified by nested PCR was described in 2016. The MLST loci had high variability, but many specimens could not be assigned a type because of poor DNA sequencing quality at one or more loci. We analyzed Cyclospora-positive stool specimens collected during 1997–2016 from 54 patients, including 51 from the United States. We noted limited inter-specimen variability for one locus (CYC15) and the frequent occurrence of unreadable DNA sequences for two loci (CYC3 and CYC13). Overall, using the remaining two loci (CYC21 and CYC22), we detected 17 different concatenated sequence types. For four of five clusters of epidemiologically linked cases for which we had specimens from >1 case-patient, the specimens associated with the same cluster had the same type. However, we also noted the same type for specimens that were geographically and temporally unrelated, indicating poor discriminatory power. Furthermore, many specimens had what appeared to be a mixture of sequence types at locus CYC22. We conclude that it may be difficult to substantially improve the performance of the MLST method because of the nucleotide repeat features of the markers, along with the frequent occurrence of mixed genotypes in Cyclospora infections.
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spelling pubmed-63546072019-02-15 Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers Hofstetter, Jessica N. Nascimento, Fernanda S. Park, Subin Casillas, Shannon Herwaldt, Barbara L. Arrowood, Michael J. Qvarnstrom, Yvonne Parasite Research Article Cyclospora cayetanensis is a human parasite transmitted via ingestion of contaminated food or water. Cases of C. cayetanensis infection acquired in the United States often go unexplained, partly because of the difficulties associated with epidemiologic investigations of such cases and the lack of genotyping methods. A Multilocus Sequence Typing (MLST) method for C. cayetanensis based on five microsatellite loci amplified by nested PCR was described in 2016. The MLST loci had high variability, but many specimens could not be assigned a type because of poor DNA sequencing quality at one or more loci. We analyzed Cyclospora-positive stool specimens collected during 1997–2016 from 54 patients, including 51 from the United States. We noted limited inter-specimen variability for one locus (CYC15) and the frequent occurrence of unreadable DNA sequences for two loci (CYC3 and CYC13). Overall, using the remaining two loci (CYC21 and CYC22), we detected 17 different concatenated sequence types. For four of five clusters of epidemiologically linked cases for which we had specimens from >1 case-patient, the specimens associated with the same cluster had the same type. However, we also noted the same type for specimens that were geographically and temporally unrelated, indicating poor discriminatory power. Furthermore, many specimens had what appeared to be a mixture of sequence types at locus CYC22. We conclude that it may be difficult to substantially improve the performance of the MLST method because of the nucleotide repeat features of the markers, along with the frequent occurrence of mixed genotypes in Cyclospora infections. EDP Sciences 2019-01-31 /pmc/articles/PMC6354607/ /pubmed/30702060 http://dx.doi.org/10.1051/parasite/2019004 Text en © J.N. Hofstetter et al., published by EDP Sciences, 2019 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Hofstetter, Jessica N.
Nascimento, Fernanda S.
Park, Subin
Casillas, Shannon
Herwaldt, Barbara L.
Arrowood, Michael J.
Qvarnstrom, Yvonne
Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers
title Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers
title_full Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers
title_fullStr Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers
title_full_unstemmed Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers
title_short Evaluation of Multilocus Sequence Typing of Cyclospora cayetanensis based on microsatellite markers
title_sort evaluation of multilocus sequence typing of cyclospora cayetanensis based on microsatellite markers
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6354607/
https://www.ncbi.nlm.nih.gov/pubmed/30702060
http://dx.doi.org/10.1051/parasite/2019004
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