食源性金黄色葡萄球菌肠毒素基因的多重PCR快速检测法

Multiplex PCR for rapid detection of enterotoxin genes in foodborne Staphylococcus aureus

目的建立食源性金黄色葡萄球菌肠毒素基因的多重PCR快速检测方法。方法根据公布的金黄色葡萄球菌肠毒素基因A～D型序列,分别设计引物并验证引物的特异性,建立多重PCR方法,检测食品风险监测和食物中毒事件中分离得到的165株金黄色葡萄球菌肠毒素基因A～D型。结果 165株金黄色葡萄球菌有95株携带肠毒素基因,毒素基因携带率为57.6%(95/165),携带一种基因型的占42.4%(72/165),同时携带两种以上毒素基因的占13.9%(23/165)。结论该方法具有简便快速、灵敏度高、特异性强等特点,适用于食源性金黄色葡萄球菌和食物中毒中金黄色葡萄球菌肠毒素的检测。

Objective To establish a multiplex polymerase chain reaction （PCR） method for screening the foodborne Staphylococcus aureus enterotoxin（SE） genes. Methods Four primers were designed according to the published Staphylococcus aureus toxin genes conserved sequence. The specificity of four primers were tested by single PCR method and sequencing. A multiplex PCR method was established and used for detecting 165 strains of Staphylococcus aureus isolated from food risk monitoring and food poisoning events. Results A total of 165 strains of Staphylococcus aureus were detected with their typeA-typeD enterotoxin genes in this study,in which 95 （57.58%） strains carried at least one kind of enterotoxin genes; 42.42%（72/165） of strains carried one enterotoxin gene type, while 13.94%（23/165） of strains carried two or more types of enterotoxin gene. Conclusion This multiplex PCR method, which has been gradually applied to foodborne Staphylococcus aureus and food poisoning events, is rapid, sensitive, specific and is applicable for the detection of Staphylococcus aureus enterotoxin genes in food risk monitoring and food poisoning diagnosis.