Submitted to: Journal of Clinical Microbiology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 10/27/2010
Publication Date: 1/1/2011
Publication URL: handle.nal.usda.gov/10113/53695
Citation: Feng, Y., Yang, W., Ryan, U., Zhang, L., Kvac, M., Fayer, R., Koudela, B. 2011. Development of a multilocus sequence typing tool for Cryptosporidium muris and Cryptosporidium andersoni. Journal of Clinical Microbiology. p. 34-41. Interpretive Summary: The two most common species of Cryptosporidium parasites found in the stomachs of mammals are C. muris and C. andersoni. Nearly 20 other species have been identified in the intestines of mammals. Natural infections with C. muris have been reported in hamsters, squirrels, Siberian chipmunks, Algerian mice, common mice, wood mice, bank voles, rock hyrax, bactrian camels, mountain goats, cynomolgus monkeys, bilbies, pigs, dogs, and humans. Natural infections with C. andersoni have been reported in cattle worldwide. Molecular based tests have been widely used to study the epidemiology of intestinal Cryptosporidium species but not the gastric species. In the present study, 4 gene sequences were chosen from 20 potential candidates for test development. From 27 C. muris and 17 C. andersoni DNA preparations 11 C. muris and 7 C. andersoni multilocus sequence typing subtypes were detected. This high resolution typing method should be useful for identifying and tracing the epidemiology of gastric Cryptosporidium species.
Technical Abstract: Although genotyping tools have been developed and widely used in the characterization of the transmission of intestinal Cryptosporidium spp., they 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 genetic loci. In all analyses, the C. muris isolate (TS03) originated from an East African mole rat (Tachyoryctes splendens) 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.