应用实时荧光PCR技术检测金黄色葡萄球菌肠毒素基因  

Application of real-time fluorescent PCR technique in detection of enterotoxin genes in Staphyloccocus aureus

作　　者：葛小萍[1] 陈棋炯[1] 孙永祥[1] 傅丹青[1] 丁水军[1] 戴城钢[1] 

机构地区：[1]浙江省杭州市萧山区疾病预防控制中心,杭州311201

出　　处：《中国卫生检验杂志》2012年第9期2099-2101,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的:建立一种简便、特异的荧光PCR检测方法,用于金黄色葡萄球菌肠毒素的检测。方法:按金黄色葡萄球菌SEA～SEE型肠毒素基因序列设计引物,在普通PCR检测体系中,加入SYBR Green I荧光染料,建立荧光PCR检测体系。结果:46株金黄色葡萄球菌中检出22株携带肠毒素基因,阳性率为47.83%。以SEA、SEB检出率较高,分别为31.82%和27.27%;不同来源的分离株携带肠毒素基因的比例不同,同时携带2种及以上毒素基因的菌株占27.27%。结论:荧光PCR检测金黄色葡萄球菌肠毒素的方法具有快速、敏感、特异性高的特点,适用于肠毒素基因的分型与分布的研究,适合基层疾控部门使用。Objective：Using real-time fluorescent PCR method to detect enterotoxin genes in Staphylococcus aureus（S.aureus） isolates from different sources.Methods： Primers were designed based on enterotoxin SEA^SEE genes sequences from S.aureus and applied with SYBR Green I fluorescent dye to establish real-time PCR system.Results： 22 strains carrying enterotoxin genes were detected from 46 S.aureus strains,and the positive rate of enterotoxin gene in isolates was 47.83%.The most frequently found genes were SEA（31.82%） and SEB（27.27%）.The detection rates of enterotoxin gene from different sources were different,and the isolates with more than one toxin gene accounted for 27.27%.Conclusion： SYBR Green I incorporation assay of real-time PCR can be applied in gene typing of S.aureus enterotoxin,which is more rapid,sensitive and specific.The typing and screening method could be promoted to use in grass roots CDC conveniently.

关 键 词：金黄色葡萄球菌 肠毒素 荧光PCR 分型 

分 类 号：R446.5[医药卫生—诊断学]