快速检测金黄色葡萄球菌环介导等温扩增方法的建立与评价

Establishment and evaluation of a rapid method for detection of Staphylococcus aureus by loop-mediated isothermal amplification

目的探讨环介导等温扩增技术(LAMP)作为快速检测方法对金黄色葡萄球菌快速检测的可行性和可靠性。方法建立LAMP反应体系,优化反应条件,进行灵敏度和特异度试验。结果筛选出了LAMP反应检测金黄色葡萄球菌nuc基因的最佳引物组,最佳反应温度为62℃,LAMP和PCR检测nuc基因的最低检测限分别为1.47、14.7pg/μL,检测金黄色葡萄球菌的特异度为100%。结论 LAMP具有操作简单、敏感度高、特异性强的特点,能够满足基层实验室、应急检测或现场检测等方面的使用需求。

Objective To investigate the feasibility and reliability of loop-mediated isothermal amplification（LAMP）in rapid detection of Staphylococcus aureus as a rapid detection method.Methods LAMP reaction system was established,the reaction conditions was optimized,the sensitivity and specificity experiments were carried out.Results The best primer set was selected to detect staphylococcus aureus for nuc genes through LAMP reaction.The best reaction temperature selected was 62 ℃,and the lowest detectable limits of LAMP and PCR for nuc genes were1.47,14.7pg/μL respectively.The specificities of two methods to detect Staphylococcus aureus were 100%.Conclusion LAMP detection method is simple,safe operation,with high sensitivity and high specificity,which can meet the need of grassroots laboratory,emergency detection and on-site testing and other aspects.