Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/15/2010
Publication Date: 1/3/2011
Citation: Feng, Y., Yang, W., Ryan, U., Zhang, L., Kvac, M., Koudela, B., Modry, D., Li, N., Fayer, R., Xiao, L. 2011. Development of a multilocus sequence typing tool for Cryptosporidium muris and Cryptosporidium andersoni. Journal of Clinical Microbiology. 49(1):34-41. Interpretive Summary: A diagnostic technique was developed based on high resolution screening of multiple genes of C. muris and C. andersoni, two species of Cryptosporidium that strongly resemble one another physically as well as both being gastric parasites of a wide range of mammals, including humans. The technique will be useful in population genetics studies that can track transmission of gastric Cryptosporidium species from a wide range of host sources and thereby determine pathways of environmental contamination.
Technical Abstract: Although widely used in the characterization of the transmission of intestinal Cryptosporidium spp., genotyping tools are not available for C. muris and C. andersoni, two most common gastric Cryptosporidium spp. of mammals. In this study, we screened the C. muris whole genome sequencing data for microsatellite and minisatellite sequences. Among the 13 potential loci (six microsatellite and seven minisatellite loci) evaluated by PCR and DNA sequencing, four were eventually chosen. DNA sequence analyses of 27 C. muris and 17 C. andersoni DNA preparations showed the presence of 5-10 subtypes of C. muris and 1-4 subtypes of C. andersoni at each locus. Altogether, 11 C. muris and seven C. andersoni multilocus sequence typing (MLST) subtypes were detected among the 16 C. muris and 12 C. andersoni specimens successfully sequenced at all four loci. In all analyses, the C. muris isolate (TS03) originated from an East African mole rat differed significantly from other C. muris isolates, approaching the extent of genetic differences between C. muris and C. andersoni. Thus, a MLST technique was developed for high resolution typing of C. muris and C. andersoni. It should be useful in the characterization of the population genetics and transmission of gastric Cryptosporidium spp.