ARS | Publication request: IDENTIFICATION OF TOXIN AND PILUS GENES IN PORCINE ESCHERICHIA COLI USING POLYMERASE CHAIN REACTION (PCR) WITH MULTIPLE PRIMER PAIRS

Submitted to: American Society for Microbiology
Publication Type: Abstract Only
Publication Acceptance Date: May 8, 1997
Publication Date: N/A

Technical Abstract: We have developed a PCR method which uses multiple primer pairs in a single reaction to detect toxin and pilus genes in E. coli. Primer pairs were designed based on the known sequences for E. coli heat-labile toxin (LT), heat-stable toxins (STaP and STb), Shiga-like toxin II variant (Stx- IIe), and the pili K99, K88, F41, 987P, and F18. These 9 virulence attributes, in various combinations, are present in enterotoxin-producing and Shiga-like toxin-producing E. coli pathogenic for swine. Primer pairs were designed to produce amplified products with unique sizes which could be distinguished by agarose gel electrophoresis. All primers had similar melting temperatures and minimal predicted interactions with other primers in the mixture. There was a high correlation between virulence genes identified using multiple primer PCR and by colony blot hybridization. This PCR procedure rapidly and accurately identifies virulence genes and may be useful for the differentiation of nonpathogenic and pathogenic E. coli.