Abstract: The present invention describes the development and evaluation of the sensitivity and specificity of a PCR-based 5′ nuclease assay for presumptively detecting E. coli O157:H7 DNA. The specificity of the eaeA-based, 5′ nuclease assay system was sufficient to correctly identify all E. coli O157:H7 strains evaluated, mirroring the previously described specificity of the PCR primers. The SZ-primed, eaeA-targeted, 5′ nuclease detection assay was capable of rapid, semiautomated, presumptive detection of E. coli O157:H7 when ≧103 CFU/ml were present in modified tryptic soy broth (mTSB) or modified E. coli broth (mEC), and when ≧104 CFU/ml were present in ground beef-mTSB mixtures. Incorporating an immunomagnetic separation step (IMS), followed by a secondary enrichment culturing step and with DNA recovery, improved the detection threshold to ≧102 CFU/ml.

Abstract: The use of specific primers and a probe in a reverse transcriptase-polymerase chain reaction provides a method for specifically identifying bovine respiratory syncytial virus in cattle.

Abstract: The use of E. suis specific primers in PCR with DNA from swine blood increases the sensitivity of current DNA hybridization protocols for determining whether swine are infected with E. suis prior to the development of any clinical symptoms. The present invention provides these E. suis primers and a method to use these primers in a PCR protocol to provide a highly sensitive diagnostic assay for early signs of an E. suis infection.