致猪水肿病大肠埃希菌多重PCR检测方法的建立及应用

doi:10.11843/j.issn.1671-7236.2015.03.005

Abstract

Abstract: The aim of this study was to establish a rapid multiplex PCR detection method for swine edema disease caused by Shiga toxin-producing Escherichia coli (STEC).Three pairs of specific primers based on Escherichia coli 16S rDNA,Shiga toxin Stx2e subunit A and fimbriae F18ab subunit A were designed and the reaction conditions were optimized.At the same time,sensitivity and specificity of multiplex PCR were studied.The results showed that the amplification product sizes of positive control strain were 1 062,733 and 313 bp.The specificity and sensitivity tests showed that there was no cross reaction with Salmonella enteritidis,Pasteurella multocida,Actinobacillus pleuropneumoniae,Haemophilus parasuis,Bordetella bronchiseptica and Streptococcus suis,as little as 1 875 CFU of bacteria could be detected by multiplex PCR.128 strains of pathogenic Escherichia coli isolated from swine were detected by multiplex PCR,36 strains of STEC were found,30 of them contained both Stx2e and F18ab fimbriae genes,and the others only had Stx2e gene.The results showed that the multiplex PCR could be used in rapid diagnosis and epidemiological investigation of swine edema disease.

Key words: swine edema disease; Escherichia coli; Shiga toxin Stx2e; F18ab fimbriae

CLC Number:

LI Wei-jie, ZHAO Yun, WEI Cai-wen, QI Xiao-xin, TIAN Ye, JIANG Tao-zhen. Establishment and Application of Multiplex PCR to Identify Escherichia coli Causing Swine Edema Disease[J]. , 2015, 42(3): 537-543.

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Establishment and Application of Multiplex PCR to Identify Escherichia coli Causing Swine Edema Disease

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