Design, development, and testing of a new multi-locus sequence typing scheme for the zoonotic pathogen Cryptosporidium parvum

The zoonotic parasite <i>Cryptosporidium parvum</i> is an important global cause of diarrheal disease in humans and young ruminants. Molecular typing is essential to track transmission routes and identify clusters of cases. Here, we developed a novel multi-locus sequence typing (MLST) scheme based on single nucleotide polymorphisms (SNPs) in unlinked markers. Coding regions with high variability were identified by comparing whole genome sequences (WGS) from 43 human- and 92 ruminant-derived <i>C. parvum</i> samples collected across Europe. We first selected 18 markers and showed that they provide high discrimination among the samples with WGS data, with 88% of the MLSTs being singletons. Next, we defined a MLST based on eight genetically unlinked markers and generated sequence data from 305 <i>C. parvum</i> samples, collected from four different host species and 13 European countries. We consolidated a set of 365 fully genotyped samples, characterized by the presence of 154 different MLSTs, 105 of which were singletons. Network analyses showed no complete clustering of samples by host species or country of origin at the European scale. We further showed that samples with <i>gp60</i> subtypes that are common in Europe are divided into many MLSTs by the new scheme, highlighting its increased discriminatory ability. However, the applicability of the scheme in public health settings is limited by its cost, turnaround time, and scalability. To achieve discrimination of <i>C. parvum</i> samples based on SNPs, a large number of loci needs to be analysed, and this is feasible using amplicon sequencing technologies.

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Publication Details

Journal
Current Research in Parasitology and Vector-Borne Diseases
Published
2025-01-01
DOI
https://doi.org/10.1016/j.crpvbd.2025.100308
Citations
1
Primary Topic
Zoonotic diseases and public health
Type
article
Field-Weighted Citation Impact
1.88
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article

Design, development, and testing of a new multi-locus sequence typing scheme for the zoonotic pathogen Cryptosporidium parvum

Guy Robinson, Christen Rune Stensvold, Emma Östlund, Martha Betson et al.
1 citations
Current Research in Parasitology and Vector-Borne Diseases
Zoonotic diseases and public health
1.88
article

Design, development, and testing of a new multi-locus sequence typing scheme for the zoonotic pathogen Cryptosporidium parvum

Guy Robinson, Christen Rune Stensvold, Emma Östlund, Martha Betson, Christian Klotz, Simone M. Cacciò, Anton de Jong, Jonas Johansson Wensman, Rachel M. Chalmers, Jeroen Roelfsema, Karim Tarik Adjou, S. Ptochos, Barbara Šoba, Ralf Ignatius, Grégory Karadjian, Karin Troell, Umer Chaudhry, Anna Rosa Sannella, Rebecca K. Davidson, Paolo Vatta, Jacek Sroka, Lauren Davies
article en
1 citations

Abstract

The zoonotic parasite <i>Cryptosporidium parvum</i> is an important global cause of diarrheal disease in humans and young ruminants. Molecular typing is essential to track transmission routes and identify clusters of cases. Here, we developed a novel multi-locus sequence typing (MLST) scheme based on single nucleotide polymorphisms (SNPs) in unlinked markers. Coding regions with high variability were identified by comparing whole genome sequences (WGS) from 43 human- and 92 ruminant-derived <i>C. parvum</i> samples collected across Europe. We first selected 18 markers and showed that they provide high discrimination among the samples with WGS data, with 88% of the MLSTs being singletons. Next, we defined a MLST based on eight genetically unlinked markers and generated sequence data from 305 <i>C. parvum</i> samples, collected from four different host species and 13 European countries. We consolidated a set of 365 fully genotyped samples, characterized by the presence of 154 different MLSTs, 105 of which were singletons. Network analyses showed no complete clustering of samples by host species or country of origin at the European scale. We further showed that samples with <i>gp60</i> subtypes that are common in Europe are divided into many MLSTs by the new scheme, highlighting its increased discriminatory ability. However, the applicability of the scheme in public health settings is limited by its cost, turnaround time, and scalability. To achieve discrimination of <i>C. parvum</i> samples based on SNPs, a large number of loci needs to be analysed, and this is feasible using amplicon sequencing technologies.

Current Research in Parasitology and Vector-Borne DiseasesVol. 8
Statens Serum Institut (DK), Long Island University (US), University of Ljubljana (SI), École Nationale Vétérinaire d'Alfort (FR), Robert Koch Institute (DE), Istituto Superiore di Sanità (IT), Singleton Hospital (GB), University of Surrey (GB), Swedish University of Agricultural Sciences (SE), Swansea University (GB), Public Health Wales (GB), Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement (FR), National Institute for Public Health and the Environment (NL), National Veterinary Research Institute (PL), Norwegian Veterinary Institute (NO), Charité - Universitätsmedizin Berlin (DE)
Openalex Percentile: Top 12%
Zoonotic diseases and public health
1.88
DOIOpen Access PDFOpenAlex
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