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Multiplex PCR for detection of virulence gene of enterotoxigenic Escherichia coli

Liandong Qu

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Abstract

To detect and identify the 2 fimbriae genes(K88,K99) and toxin genes of heat-stable enterotoxin a(STa) in enterotoxigenic Escherichia coli(ETEC),a multiplex PCR method was developed based on 3 pairs of primers for K88,K99 and STa gene amplification.The reference E.coli strains which passed the different virulence genes were detected and the results demonstrated that 237 bp,314 bp and 166 bp DNA fragments of K88,K99 and STa genes were amplified by this protocol,respectively.The multiplex PCR was proved to be specificity and sensitivity.Furthermore,a total of 23 E.coli were tested and the results showed 2 samples were K99/STa and 1 sample was STa positive.The multiplex PCR assay provides a method to detect ETEC which caused diarrhea in piglets.

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What this paper is about

To detect and identify the 2 fimbriae genes(K88,K99) and toxin genes of heat-stable enterotoxin a(STa) in enterotoxigenic Escherichia coli(ETEC),a multiplex PCR method was developed based on 3 pairs of primers for K88,K99 and STa gene amplification.The reference E.coli strains which passed the different virulence genes were detected and the results demonstrated that 237 bp,314 bp and 166 bp DNA fragments of K88,K99 and STa genes were amplified by this protocol,respectively.The multiplex PCR was proved to be specificity and sensitivity.Furthermore,a total of 23 E.coli were tested and the results showed 2 samples were K99/STa and 1 sample was STa positive.The multiplex PCR assay provides a method to detect ETEC which caused diarrhea in piglets.

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Available abstract

To detect and identify the 2 fimbriae genes(K88,K99) and toxin genes of heat-stable enterotoxin a(STa) in enterotoxigenic Escherichia coli(ETEC),a multiplex PCR method was developed based on 3 pairs of primers for K88,K99 and STa gene amplification.The reference E.coli strains which passed the different virulence genes were detected and the results demonstrated that 237 bp,314 bp and 166 bp DNA fragments of K88,K99 and STa genes were amplified by this protocol,respectively.The multiplex PCR was proved to be specificity and sensitivity.Furthermore,a total of 23 E.coli were tested and the results showed 2 samples were K99/STa and 1 sample was STa positive.The multiplex PCR assay provides a method to detect ETEC which caused diarrhea in piglets.

Key concepts: Enterotoxigenic Escherichia coli, Multiplex polymerase chain reaction, Biology, Virulence, Escherichia coli, Enterotoxin, Microbiology, Gene

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