实时荧光定量PCR检测金黄色葡萄球菌方法的试验研究  

Real-time fluorescence quantitative PCR detection of Staphylococcus aureus in experimental study

作　　者：苏明权[1] 杨柳[1] 马越云[1] 常亮[1] 肖凤静[1] 郝晓柯[1] 

机构地区：[1]第四军医大学附属西京医院全军临床检验医学中心,西安710032

出　　处：《国际检验医学杂志》2010年第8期794-796,共3页International Journal of Laboratory Medicine

摘　　要：目的建立金黄色葡萄球菌实时荧光定量PCR的快速检测方法,探讨该方法的可行性和应用价值。方法根据金黄色葡萄球菌femB基因序列设计引物和探针,采用基因重组技术构建用于金黄色葡萄球菌检测的定量标准品,建立实时荧光定量PCR检测金黄色葡萄球菌的方法。结果成功构建了金黄色葡萄球菌重组质粒标准品和金黄色葡萄球菌实时荧光定量PCR方法;通过特异性、敏感性、稳定性和重复性试验,结果表明具有较好的特异性、敏感性、稳定性和重复性;将模拟标本与分离培养对比,两者符合率为100%。结论金黄色葡萄球菌实时荧光定量PCR检测方法的建立,为金黄色葡萄球菌感染诊断及食源性金黄色葡萄球菌污染的快速检测提供依据,可用于临床感染诊断及食品卫生监管、商品检验检疫等。Objective To establish real-time fluorescent quantitative PCR method for rapid detecting Staphylococcus aureus and investigate its feasibility and application value. Methods Primers and probes were designed based on femB gene sequence of Staphylococcus aureus. Quantitative standard for Staphylococcus aureus was upbuih by cloning of femg gene. Then real-time fluo- rescence quantitative PCR for Staphylococcus aureus was established routinely. Results It was identified that femB gene fragment was cloned as standard successfully. Real time fluorescence quantitative PCR method showed good specificity, sensitivity, stability by the verification experiment. And the coincidence rate was 100% comparing simulation specimens with culture strains. Conclusion The establishment of real-time fluorescent quantitative PCR detection of Staphylococcus aureus provide a basis for rapid detecting food-borne contamination of Staphylococcus aureus. It could be applied to food hygiene regulation,quarantine and clinical diagnosis.

关 键 词：葡萄球菌 金黄色 聚合酶链反应 葡萄球菌感染 诊断 

分 类 号：R440[医药卫生—诊断学]