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赵爱兰, 熊衍文, 白雪梅, 张少敏, 王艳, 孙晖, 叶长芸. 鉴定五类致泻性大肠埃希菌和志贺菌的多重PCR方法[J]. 疾病监测, 2011, 26(1): 65-67.
DOI:
10.3784/j.issn.1003-9961.2011.01.020
Citation: ZHAO Ai-lan, XIONG Yan-wen, BAI Xue-mei, ZHANG Shao-min, WANG Yan, SUN Hui and YE Chang-yun. Multiplex PCR for identification of diarrheogenic Escherichia coli and Shigella spp[J]. Disease Surveillance, 2011, 26(1): 65-67. DOI: 10.3784/j.issn.1003-9961.2011.01.020
Citation: ZHAO Ai-lan, XIONG Yan-wen, BAI Xue-mei, ZHANG Shao-min, WANG Yan, SUN Hui and YE Chang-yun. Multiplex PCR for identification of diarrheogenic Escherichia coli and Shigella spp[J]. Disease Surveillance, 2011, 26(1): 65-67. DOI: 10.3784/j.issn.1003-9961.2011.01.020
鉴定五类致泻性大肠埃希菌和志贺菌的多重PCR方法
摘要:
目的 建立一种快速鉴定和鉴别五类致泻性大肠埃希菌和志贺菌的多重PCR方法。
方法 利用五类致泻性大肠埃希菌和志贺菌的毒力基因stx1、stx2、eaeA、lt、stIb、aggR、ipaH及16S rRNA基因rrs,建立8重PCR反应体系,并对PCR引物、反应体系和循环条件进行优化。
结果 8对PCR引物均能特异地扩增相应的基因。对本实验室87株大肠埃希菌及志贺菌的检测结果与先前应用单重PCR的鉴定结果完全一致。
结论 采用内对照基因和毒力基因建立的多重PCR方法既能在一个反应体系中鉴定和区分五类致泻性大肠埃希菌和志贺菌,又能评价PCR体系的反应效率,为五类致泻性大肠埃希菌的快速鉴定和鉴别提供了一种有效检测手段。
English
Multiplex PCR for identification of diarrheogenic Escherichia coli and Shigella spp
Abstract:
Objective To establish a rapid multiplex PCR assay to identify and differentiate the five main pathotypes of diarrheogenic Escherichia coli and Shigella spp.
Methods The virulence genes stx1, stx2, eaeA, lt, stIb, aggR, ipaH as the target genes and 16S RNA gene rrs as an internal control gene were grouped into a multiplex PCR reaction system. By optimizing the multiplex PCR primers, reaction conditions and cycle parameters, the multiplex PCR can identify and differentiate the five main pathotypes of diarrheogenic Escherichia coli and Shigella spp rapidly and accurately.
Results The established multiplex PCR assay was applied in this study to identify 87 strains of Escherichia coli and Shigella spp. The results showed that the genetypes were 100% consistent with those of single PCR amplification.
Conclusion The multiplex PCR assay in this study can be used not only to identify the five main pathotypes of diarrheogenic Escherichia coli and Shigella spp in a single reaction, but also to evaluate the efficiency. This assay is an effective supplement to other molecular Methods in the identification of the five main pathotypes of diarrheogenic Escherichia coli and Shigella spp.
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Key words:
- diarrheogenic Escherichia coli /
- multiplex PCR /
- internal control gene
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