Modification of a rapid method for the identification of gene-specific polymorphisms in Cryptosporidium parvum and its application to clinical and epidemiological investigations - PubMed (original) (raw)

Modification of a rapid method for the identification of gene-specific polymorphisms in Cryptosporidium parvum and its application to clinical and epidemiological investigations

K Elwin et al. Appl Environ Microbiol. 2001 Dec.

Abstract

The application of genotyping to clinical isolates of Cryptosporidium has increased significantly our knowledge and understanding of the distribution and epidemiology of this parasite. However, some methods can be laborious and demand specialist technical expertise. PCR-restriction fragment length polymorphism (RFLP) techniques represent a more rapid and simple method of genotyping to support epidemiological and clinical investigations than conventional DNA analytical techniques. We describe a nested PCR-RFLP technique that identifies polymorphisms in the C. parvum thrombospondin-related adhesive protein gene locus; this method offers a sensitive and specific tool for the confirmation and investigation of disease associated with C. parvum. The potential of this enhanced method is demonstrated by its application to the confirmation and epidemiological investigation of an outbreak of cryptosporidiosis associated with a school visit to an open farm.

PubMed Disclaimer

Figures

FIG. 1

FIG. 1

Agarose gel electrophoresis of _Bst_EII and _Hae_III restriction endonuclease cleavage products. Lanes 1 and 11, DNA molecular-weight-standard marker. Lane 2, C. parvum (Moredun strain, genotype 2) _Bst_EII cleavage products. Lane 3, C. parvum (Moredun strain, genotype 2) _Hae_III-resistant PCR product. Lane 4, C. parvum (IOWA strain, genotype 2) _Bst_EII cleavage products. Lane 5, C. parvum (IOWA strain, genotype 2) _Hae_III-resistant product. Lane 6, C. parvum (human isolate, genotype 1) _Bst_EII-resistant product. Lane 7, C. parvum (human isolate, genotype 1) _Hae_III cleavage products. Lane 8, C. parvum (human isolate, genotype 2) _Bst_EII cleavage products. Lane 9, C. parvum (human isolate, genotype 2) _Hae_III-resistant product. Lane 10, PCR negative control.

References

    1. Caccio S, Homan W, Camilli R, Traldi G, Kortbeek T, Pozio E. A microsatellite marker reveals population heterogeneity within human and animal genotypes of Cryptosporidium parvum. Parasitology. 1999;120:237–244. - PubMed
    1. Carraway M, Tzipori S, Widmer G. New RFLP marker in Cryptosporidium parvum identifies mixed parasite populations and genotypic instability in response to host change. Infect Immun. 1997;65:3958–3960. - PMC - PubMed
    1. Casemore D P. Human cryptosporidiosis: clinical aspects, epidemiology and control. Proc R Coll Physicians (Edinburgh) 2000;30:287–293.
    1. Chalmers R M, Elwin K. Implication and importance of genotyping Cryptosporidium. Communicable Dis Public Health. 2000;3:155–157. - PubMed
    1. Clark D P. New insights into human cryptosporidiosis. Clin Microbiol Rev. 1999;12:554–563. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources