多重PCR检测金黄色葡萄球菌六型肠毒素基因的研究

Study on fast detection of six kinds of Staphylococcus aureus enterotoxins genes with a multiplex PCR method

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、478、257、319、170、375bp的目的片段,表明6对引物具有良好的特异性。结论: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。

Objective： To establish a multiplex PCR method for detecting SEA, SEB, SEC, SED, SEE and SEH genes of Staphylococcus aureus fast and highly effectively. Methods： According to SEA, SEB, SEC, SED, SEE and SEH gene sequences of Staphylococcus 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, 478, 257, 319, 170, 375 bp were simultaneously amplified from six couple primers. The results showed that the PCR of six pairs primers were good specifity. Condusions： A simple, rapid and sensitive multiplex PCR method for detecting six kinds of S. aureus enterotoxin genes has been established. It could detect SEA, SEB, SEC, SED, SEE and SEH genes simultaneously, therefore, the prospect of fast detection of Staphylococcus aureus enterotoxin is brilliant.