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Applied and Environmental Microbiology, December 2001, p. 5581-5584, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5581-5584.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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,1 R. Roberts,2 E. C. Guy,1 and D. P. Casemore3

PHLS Cryptosporidium Reference Unit, Swansea Public Health Laboratory, Singleton Hospital, Swansea, United Kingdom SA2 8QA1; North Wales Health Authority, Preswylfa, Mold, United Kingdom CH7 1PZ2; and Rhyl Public Health Laboratory, Glan Clwyd District General Hospital, Rhyl, Denbighshire, United Kingdom LL18 5UJ3

Received 24 May 2001/Accepted 15 August 2001

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.

* Corresponding author. Mailing address: PHLS Cryptosporidium Reference Unit, Swansea Public Health Laboratory, Singleton Hospital, Swansea, United Kingdom SA2 8QA. Phone: 0044 (0)1792 285055. Fax: 0044 (0)1792 202320. E-mail: kristin.elwin{at}phls.wales.nhs.uk.

Applied and Environmental Microbiology, December 2001, p. 5581-5584, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5581-5584.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.