多重PCR 检测金黄色葡萄球菌六型肠毒素基因的研究.doc

文档介绍：多重PCR检测金黄色葡萄球菌六型肠毒素基因的研究
刘继超,姜铁民,姜阿赤,陈历俊通讯作者: 陈历俊,chlj@
基金项目: “十一五”国家科技支撑计划(2009BADB9B06);北京市重点科技计划项目(D10**********)
作者简介: 刘继超,男,1983年出生,硕士,乳品微生物。
(北京三元食品股份有限公司,北京 100076)
摘要:目的:为了建立一种简单、快速检测金黄色葡萄球菌六型肠毒素基因的多重PCR方法。方法:根据相关文献和Genebank报道的编码金黄色葡萄球菌肠毒素A、B、C、D、E、H的基因序列,选择合成了6对特异性引物,建立多重PCR体系,并对反应条件进行了优化。结果:6对引物能同时特异地扩增出120 bp、478 bp、257 bp、319 bp、170 bp和375 bp的目的片段,表明6对引物具有良好的特异性。结论:本研究成功地建立了一种同时检测金黄色葡萄球菌六型肠毒素基因的多重PCR方法,在金黄色葡萄球菌肠毒素快速筛查方面具有良好的应用前景。
关键词:多重PCR;检测;金黄色葡萄球菌;肠毒素
A study on fast detection of six kinds of us aureus enterotoxins genes with a multiplex PCR method
LIU Ji-chao, JIANG Tie-min, JIANG A-chi, CHEN Li-jun *
(Beijing Sanyuan Foods ., Beijing, 100076)
Abstract: Objective: To establish a multiplex PCR method for detecting SEA, SEB, SEC, SED, SEE and SEH genes of us aureus fast and highly effectively. Methods: According to SEA, SEB, SEC, SED, SEE and SEH gene sequences of us aureus in GenBank and relevant literatures, six pairs of specific primers had been designed. Established multiplex PCR methods and its reaction conditions were optimized. Results: Under the optimized conditions, the six fragments of 120 bp, 478 bp, 257 bp, 319 bp, 170 bp and 375 bp were simultaneously amplified from six couple primers. The results showed that the PCR of six pairs primers were good specifity. Conclusions: A simple, rapid and sensitive multiplex PCR method for detecting