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United States Department of Agriculture

Agricultural Research Service

Title: Application of a Multiplex Pcr Assay for Simultaneous Detection of Shiga Toxigenic Escherichia Coli and Enterohemorrhagic E. Coli of Serotypes O157:h7, O26, and O111

item Sharma, Vijay
item Casey, Thomas

Submitted to: International Symposium and Workshop on Shiga Toxin ... Escherichia coli
Publication Type: Abstract Only
Publication Acceptance Date: November 2, 2000
Publication Date: N/A

Technical Abstract: Enterohemorrhagic E. coli (EHEC) causes a broad spectrum of diseases, ranging from diarrhea to hemolytic syndrome, in humans. While most outbreaks in North America have been attributed to E. coli O157:H7, EHEC isolates belonging to serotypes O26 and O111 are emerging as important pathogens throughout the world. Since cattle are an important reservoir for EHEC, fecal contamination of meats is an important source for transmission of these pathogens to humans. Rapid and accurate diagnostic tests for detecting multiple EHEC serotypes are needed for monitoring the prevalence of these serotypes in cattle and foods of bovine origin. Moreover, these tests will also find use in estimating the frequency with which these strains are recovered from patients exhibiting symptoms of hemorrhagic colitis and hemolytic syndrome. The present study was undertaken to develop and evaluate a multiplex PCR for rapid detection of EHEC serotypes O157:H7, O26, and O111 and to differentiate these serotypes from other Shiga toxin-producing E. coli (STEC). A total of five primer pairs were used in this PCR. Three of the five primer pairs were designed to amplify a portion of the eae gene, allowing the detection of three EHEC serotypes. The remaining two primer pairs enabled the amplification and detection of stx1 and stx2, the genes encoding Shiga toxin 1 and 2, respectively, of STEC. We are currently evaluating the sensitivity of this PCR to detect EHEC and STEC in meats and feces using agarose gel- and TaqMan-based detection platforms.

Last Modified: 4/22/2015
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