Simplified method for recovery and PCR detection of Cryptosporidium DNA from bovine feces.

Applied and Environmental Microbiology
X LengR D Oberst

Abstract

An assay to identify Cryptosporidium DNA in bovine feces has been developed emphasizing standardization of sample preparation and simplification of the DNA recovery process for PCR amplification and DNA hybridization detection. The Cryptosporidium DNA recovery-PCR detection procedure (CR-PCR) can recover DNA suitable for PCR amplification without using or generating hazardous chemicals or wastes. In comparisons with a commercial enzyme-linked immunoassay (Color Vue-Cryptosporidium; Seradyn, Indianapolis, Ind.), the CR-PCR could detect 10(3) to 10(4) times fewer purified oocysts diluted in solution (water or buffered saline) and 10(2) times fewer oocysts from diarrheic feces and showed earlier detectability from solid, nondiarrheic feces in an experimental infection. This assay may prove useful for detecting Cryptosporidium oocysts in feces and in clarifying the role of livestock in waterborne outbreaks of cryptosporidiosis.

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Citations

Mar 7, 2001·Applied and Environmental Microbiology·J F Perz, S M Le Blancq
Apr 1, 2008·The Journal of Parasitology·Bing-Mu HsuChih-Li Lilian Hsu
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Apr 18, 1998·Applied and Environmental Microbiology·G Pereira M dasR B Lefebvre
Oct 9, 1999·Clinical Microbiology Reviews·D P Clark
Nov 5, 1997·Applied and Environmental Microbiology·I G Wilson

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