|Lewis, Earl - NOAA|
|Graczyk, Thaddeus - JHUSHPH|
Submitted to: American Association of Veterinary Parasitologists Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: July 11, 1999
Publication Date: July 11, 1999
Technical Abstract: A number of publications describe the use of Cryptosporidium parvum DNA sequences as targets for PCR-based detection protocols. Species-specific detection is usually achieved through the digestion of PCR amplicons with restriction endonucleases to generate RFLP profiles. We investigated use of a nested PCR protocol, targeting the C. parvum Cp11 ribosomal protein gene, for detection of oocysts in surface water, oysters, and bovine feces The nested PCR assay successfully detected oocysts in all these samples, and was more sensitive thatn conventional immunoflourescence microscopy and primary PCR, but required as templte purified oocysts or genomic DNA for optimum performance. Data will be presented on our efforts to make the assay species-specific through the use of oligonucleotide probes, and on the efficacy of rapid sample preparation techniques.