反转录-环介导等温扩增（Reverse transcriptase loop-mediated isothermal amplification RT-LAMP）技术是以RNA为检测模板的扩增反应。针对金黄色葡萄球菌特异保守nuc基因设计多组引物，通过对引物筛选和反应条件优化，建立了检测金黄色葡萄球菌的实时荧光RT-LAMP方法。利用该方法对菌株特异性、纯培养金黄色葡萄球菌灵敏度和人工污染脱脂乳样品灵敏度进行检测。等温65℃条件下，30min内完成RT-LAMP反应。该方法具有较强的菌株特异性，对纯培养金黄色葡萄球菌灵敏度检测限为20 fg/μL，在人工污染脱脂乳样品的检测中，灵敏度达到230 CFU/g。建立的实时荧光RT-LAMP方法可以为金黄色葡萄球菌现场快速检测提供有力手段。

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is an amplification reaction using RNA as a template for detection. A real-time fluorescent RT-LAMP method for the detection ofStaphylococcus aureuswas established by optimizing primers designed for specific conserved nuc gene and optimizing the reaction conditions. The strain-specific and the sensitivity of pure culture ofStaphylococcus aureusand artificial contamination of skim milk samples were detected by this method. The RT-LAMP amplification reaction was completed at 65°C for 30minutes. The method had a strong strain specificity, the detection limit of pureStaphylococcus aureuswas 20 fg/μL and the sensitivity was 230 CFU/g in the detection of artificial contaminated skim milk samples. The established real-time fluorescent RT-LAMP method can provide a powerful means for rapid detection ofStaphylococcus aureusin the field.