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目的建立一种简单、快速、准确、特异性强的能定量检测金黄色葡萄球菌的方法。方法以金黄色葡萄球菌FomB基因为靶序列,设计并合成引物和Taqman探针,优化引物与探针比例,调整镁离子浓度,对金黄色葡萄球菌进行荧光定量检测。结果引物与Taqman探针比例为1∶4,镁离子为2.5mmol/L时本底最低,荧光信号最强;该法的灵敏度1.0×103拷贝,能特异区分金色葡萄球菌与其他葡萄球菌。结论使用Taqman探针技术的荧光定量聚合酶链反应能够对金黄色葡萄球菌进行准确快速及定量检测。
Objective To establish a simple, rapid, accurate and specific method for the quantitative detection of Staphylococcus aureus. Methods The FomB gene of Staphylococcus aureus was used as target sequence to design and synthesize primers and Taqman probes. The ratio of primers and probes was optimized. The concentration of magnesium ions was adjusted and the fluorescence quantitative detection of Staphylococcus aureus was carried out. Results The ratio of primer to Taqman probe was 1: 4. When the concentration of magnesium ion was 2.5 mmol / L, the fluorescence signal was the strongest. The sensitivity of this method was 1.0 × 103 copies, which could distinguish Staphylococcus aureus from other staphylococci. Conclusion The fluorescence quantitative polymerase chain reaction using Taqman probe technology can detect Staphylococcus aureus accurately and rapidly and quantitatively.