SIMPLIFIED METHOD FOR RECOVERY AND PCR DETECTION OF CRYPTOSPORIDIUM DNA FROM BOVINE FECES

An assay to identify Cryptosporidium DNA in bovine feces has been deve loped emphasizing standardization of sample preparation and simplifica tion of the DNA recovery process for PCR amplification and DNA hybridi zation detection. The Cryptosporidium DNA recovery-PCR detection proce dure (CR-PCR) can recover DNA suitable for PCR amplification without u sing or generating hazardous chemicals or wastes. In comparisons with a commercial enzyme-linked immunoassay (Color Vue-Cryptosporidium; Ser adyn, Indianapolis, Ind.), the CR-PCR could detect 10(3) to 10(4) time s fewer purified oocysts diluted in solution (water or buffered saline ) and 10(2) times fewer oocysts from diarrheic feces and showed earlie r detectability from solid, nondiarrheic feces in an experimental infe ction. This assay may prove useful for detecting Cryptosporidium oocys ts in feces and in clarifying the role of livestock in waterborne outb reaks of cryptosporidiosis.