多重PCR检测13种金黄色葡萄球菌肠毒素基因方法的建立及应用

Establishment and application of multiple PCR for detecting 13 enterotoxin genes of Staphylococcus aureus

目的建立快速检测13种金黄色葡萄球菌肠毒素(Staphylococcusal enterotoxin,SE)的多重PCR方法,了解食源性SE的分布状况。方法设计13种SE基因的引物,建立单重PCR方法,测序验证引物特异性;优化多重PCR方法,检测144株食源性金黄色葡萄球菌的13种SE基因。结果建立的多重PCR方法特异、高效,13种SE基因在144株食源性金黄色葡萄球菌中的检出率由高到低依次为SEA(39.58%)、SEU(25.69%)、SEG(22.22%)、SEB(21.52%)、SEM(20.83%)、SEK(20.14%)SEQ(18.06%)、SEH(15.97%)、SEC(13.89%)、SEN(10.42%)、SEJ(8.33%)、SED(8.33%)、SEL(5.56%);65.97%的菌株至少含有13种SE基因中的1种,45.14%的菌株含有2种或2种以上SE基因。结论建立的多重PCR方法特异性高,快速简便,可用于SE基因的检测。

Objective To establish a multiple PCR method for detecting 13 Staphylococcus enterotoxin SE genes,and study the distribution of 13 enterotoxin genes in foodborne Staphylococcus aureus. Methods Thirteen pairs of primer were designed for detecting SE genes. The specificities of thirteen primers were tested by single PCR method. The multiple PCR method were established to detect 13 SE genes in 144 foodborne Staphylococcus aureus strains. Results The multiple PCR method was specific and efficient. The detection rate of 13 SE genes in 144 foodborne Staphylococcus aureus strains was in descending order for SEA（39.58%）,SEU（25.69%）,SEG（22.22%）,SEB（21.52%）,SEM（20.83%）,SEK（20.14%）,SEQ（18.06%）,SEH（15.97%）,SEC（13.89%）,SEN（10.42%）,SEJ（8.33%）,SED（8.33%）,SEL（5.56%）. 65.97% of the strains contained at least one gene of the 13 SE genes,45.14% of the strains contained two or more SE genes. Conclusion The multiple PCR method was highly specific,simple and convenient. It can be used for the scanning of the SE genes in Staphylococcus aureus.