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. 2001 Dec;67(12):5581-4.
doi: 10.1128/AEM.67.12.5581-5584.2001.

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  1 , R M Chalmers, R Roberts, E C Guy, D P Casemore
Affiliations

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.

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Figures

FIG. 1
FIG. 1
Agarose gel electrophoresis of BstEII and HaeIII restriction endonuclease cleavage products. Lanes 1 and 11, DNA molecular-weight-standard marker. Lane 2, C. parvum (Moredun strain, genotype 2) BstEII cleavage products. Lane 3, C. parvum (Moredun strain, genotype 2) HaeIII-resistant PCR product. Lane 4, C. parvum (IOWA strain, genotype 2) BstEII cleavage products. Lane 5, C. parvum (IOWA strain, genotype 2) HaeIII-resistant product. Lane 6, C. parvum (human isolate, genotype 1) BstEII-resistant product. Lane 7, C. parvum (human isolate, genotype 1) HaeIII cleavage products. Lane 8, C. parvum (human isolate, genotype 2) BstEII cleavage products. Lane 9, C. parvum (human isolate, genotype 2) HaeIII-resistant product. Lane 10, PCR negative control.
See this image and copyright information in PMC

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