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目的建立基于环介导等温扩增(LAMP)技术进行快速鉴定耐甲氧西林金黄色葡萄球菌(MRSA)的方法。方法采集自鼻拭子样品的MRSA菌株,针对MRSA耐药因子mec A所在基因组岛SCCmec,挑选具有高保守性的边界末端开放阅读框,设计特异性引物,进行LAMP快速检测,优化和确定LAMP反应体系和条件。结果以PCR法作为对照,LAMP检测MRSA的灵敏度为8×10~3 CFU/ml,PCR的灵敏度为8×10~5 CFU/ml。同时,LAMP快速检测法的检出率与特异性分别达92.7%与100.0%。结论 LAMP是可应用于MRSA检测的灵敏和特异性方法。
Objective To establish a rapid method for the identification of methicillin - resistant Staphylococcus aureus (MRSA) based on the ring - mediated isothermal amplification (LAMP) technique. Methods MRSA strains from nasal swab samples were collected. Based on the genomic island SCCmec where MRSA resistance gene mec A was located, a highly conserved border-end open reading frame was selected and specific primers were designed for rapid detection of LAMP to optimize and determine the LAMP response System and conditions. Results The sensitivity of LAMP to MRSA was 8 × 10 ~ 3 CFU / ml and the sensitivity of PCR was 8 × 10 ~ 5 CFU / ml. At the same time, the detection rate and specificity of LAMP rapid detection method were 92.7% and 100.0% respectively. Conclusion LAMP is a sensitive and specific method that can be applied to MRSA detection.