多重实时荧光定量聚合酶链反应检测携带耐药基因及肠毒素A基因的金黄色葡萄球菌
Multiplex fluorescent real-time PCR for detection of methicillin-resistance gene and enterotoxin gene A in Staphylococcus aureus
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摘要: 目的 建立一种快速鉴定携带耐药基因和肠毒素A基因的金黄色葡萄球菌的方法。 方法 根据金黄色葡萄球菌耐热核酸酶基因nuc、甲氧西林耐药基因mecA和肠毒素A基因sea的序列,设计特异性的引物和不同荧光素标记的TaqMan探针,通过反应体系和反应条件的优化,建立了能同时检测鉴定耐甲氧西林和产肠毒素A 的金黄色葡萄球菌的多重荧光PCR方法。 结果 该方法特异性好,可在同一个PCR反应体系中同时检测金黄色葡萄球菌的3种基因成分,检测效率高。灵敏度可以达到210-7g/l。 结论 该方法可用于食品样品的检测和流行病学分离株的快速鉴定。Abstract: Objective To establish a multiplex fluorescent real-time PCR assay for the detection of methicillin-resistance gene and enterotoxin gene A of Staphylococcus aureus. Methods We designed the specific primers and TaqMan probes targeting sea, mecA and nuc gene of Staphylococcus aureus. We used different fluorescents to label the probes. After optimization of reaction condition, we set up a multiplex fluorescent real-time PCR assay to detect Staphylococcus aureus rapidly and accurately. Results The result demonstrated that this assay was specific, which can identify three kinds of genes from Staphylococcus aureus in one reaction. The lowest detection limit was 210-7g/l. Conclusion This assay can be used in food sample detection and rapid identification of Staphylococcus aureus.
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