ULTRASENSITIVE DETECTION OF PATHOGENIC MICROBES

摘要

The present invention describes a 5' nuclease real-time polymerase chain reaction (PCR) approach for the quantification of total coliforms, E. coli, toxigenic E. coli O157:H7, toxigenic M. aeruginosa (microcystin hepatotoxins), Giardia lamblia , and Cryptosporidium parvum, based on the specific identified primer and probe sequences from the lacZ (E. coli), eaeA (E. coli O157:H7), mcyA (M aeruginosa), .beta. - giardin (G. lamblia), and COWP (C. parvum) genes respectively. The invention allows for the detection of all of the aforementioned microbes, with or without culture enrichments, utilizing a 5' nuclease PCR approach. The invention also provides primer and probe sequences useful to produce detectable amplicons, by any amplification metho d, which are diagnostic for such organisms.