葡萄球菌肠毒素B基因的PCR快速检测法  

Rapid detection of gene encoding staphylococcal enterotoxin B by PCR

作　　者：张勇[1] 石晓路[2] 梁伟[1] 李波[1] 陈润莉[1] 曾华书[1] 

机构地区：[1]广东省深圳市福田区疾病预防控制中心检验科,518040 [2]深圳市疾病预防控制中心

出　　处：《职业与健康》2013年第5期552-554,共3页Occupation and Health

摘　　要：目的建立快速检测葡萄球菌肠毒素B(staphylococcal enterotoxin B,SEB)基因的聚合酶链反应(PCR)方法。方法①根据SEB基因的序列,设计PCR引物特异性扩增靶基因片段。通过对1株产SEB金黄色葡萄球菌和9株对照菌株进行PCR检测,评价该方法的特异性;通过对产SEB金黄色葡萄球菌菌株做10倍系列稀释后进行PCR检测,评价该方法的敏感性;②分析30株金黄色葡萄球菌SEB基因的携带情况。结果①建立PCR方法快速检测SEB基因,扩增产物长度为494 bp。PCR反应体系中有26 CFU的产SEB金黄色葡萄球菌即可检出SEB基因;②30株分离的金黄色葡萄球菌中有2株携带有SEB基因。结论 PCR法可以快速、敏感地检测SEB基因,为金黄色葡萄球菌食物中毒诊断提供依据。[ Objective] To establish a PCR assay for the rapid and sensitive detection of Staphylococcal enterotoxin B （ SEB ） gene. [ Methods ] ①According to the sequences of the SEB gene, primers were selected and were used to amplify SEB gene. To evaluate the specificity of the assay, 1 strain of SEB positive Staphylococcus aureus and 9 strains of non-Staphylococcus aureus were tested by PCR. To evaluate the sensitivity of the assay, the strain of SEB positive Staphylococcus aureus was 10-fold serially diluted and was amplified by PCR. ②The SEB gene in 30 strains of Staphylococcus aureus were detected by PCR. [ Results] ①The PCR assay for the rapid and sensitive detection of SEB gene was well established. The length of amplification was 494 bp. The detection limit for SEB gene was 26 CFU;② Among 30 strains of Staphylococcus aureus, 2 strains were positive for SEB gene. [ Conclusion] The PCR assay can rapidly and exactly detect the SEB gene, and can provide evidence for the rapid diagnosis of Staphylococcal food poi-soning.

关 键 词：金黄色葡萄球菌 葡萄球菌肠毒素B 聚合酶链反应 

分 类 号：R446.5[医药卫生—诊断学]