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

• Published in: Applied and Environmental Microbiology 1996-02, Vol.62 (2), p.643-647
• Main Authors: Leng, X, Mosier, D A, Oberst, R D
• Format: Article
• Language: English
• Subjects: adn; amplification chaine polymerase; Animals; Bacteria; Base Sequence; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; bovin; Cattle; Cattle Diseases - diagnosis; Cattle Diseases - parasitology; Cellular biology; Cryptosporidiosis - epidemiology; Cryptosporidiosis - transmission; Cryptosporidiosis - veterinary; Cryptosporidium; Cryptosporidium - genetics; Cryptosporidium - isolation & purification; cryptosporidium parvum; Deoxyribonucleic acid; Disease Outbreaks; DNA; dna hybridization; DNA Primers - genetics; DNA, Protozoan - genetics; DNA, Protozoan - isolation & purification; faeces; feces; Feces - parasitology; Fundamental and applied biological sciences. Psychology; ganado bovino; Genetics; heces; hibridacion de adn; Humans; hybridation d' adn; immunoenzyme techniques; Manures; Mission oriented research; Molecular Sequence Data; polymerase chain reaction; Polymerase Chain Reaction - methods; reaccion de cadenas de polimerasa; technique immunoenzymatique; tecnicas inmunoenzimaticas; Water - parasitology
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.62.2.643-647.1996

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.