Construction of a Multiplex PCR Method for Detection Enterotoxigenic Escherichia coli in Fresh Meat

被引：0

作者：

Liu B. ^{[1
]}

Wang T. ^{[1
]}

Wang R. ^{[1
]}

Du S. ^{[1
]}

Lü X. ^{[1
]}

Zhang J. ^{[1
]}

机构：

[1] College of Food Science and Engineering, Northwest A&F University, Yangling, 712100, Shaanxi

来源：

Journal of Chinese Institute of Food Science and Technology | 2018年 / 18卷 / 12期

关键词：

Enterotoxigenic Escherichia coli; Fresh meat; Multiplex PCR; Rapid detection;

D O I：

10.16429/j.1009-7848.2018.12.029

中图分类号：

学科分类号：

摘要：

Escherichia coli and its pathogenic strains are the main pollution strains in fresh meat. The detection and control of these bacteria is the key point to ensure the quality of meat. This study intends to establish a rapid detection method based on selective enrichment and multiplex PCR for Eenterotoxigenic Escherichia coli in fresh meat. The E. coli-specific uidA gene, ETEC heat-labile enterotoxin gene lt and heat-stable enterotoxin gene sta were the three target genes of PCR. Through selection of specific primers, ETEC selective enrichment and separation, and PCR reaction condition optimization study, ETEC was detected from different pork. The results revealed that the stability and specificity of multiplex PCR were the best for the 25 μL reaction system when the annealing temperature was 60℃ and the primer addition amount was 0.8 μL(10 μmol/μL). After LST broth enrichment and MAC plate separation, the minimum detection limit of this method is 4 CFU/10 g. This method is simple, stable and specificity, can be used for rapid detection of ETEC in fresh meat. © 2018, Editorial Office of Journal of CIFST. All right reserved.

引用

页码：225 / 231

页数：6

共 10 条

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