摘要：目的：建立环介导等温扩增（LAMP）技术快速检测临床标本中金黄色葡萄球菌(SA)的方法。 方法：基于金黄色葡萄球菌femA基因部分序列设计一套（共4条）特异性引物，包括特异性识别靶序列上6个不同区域的两条内引物和两条外引物。通过条件优化，建立检测SA的LAMP方法。用该法检测临床分离的40株SA、12种其他革兰阳性球菌和8种革兰阴性杆菌，对方法特异性进行评价。SA标准菌株ATCC 25923进行10n稀释后测定方法的灵敏度。用LAMP方法及普通培养法检测临床60份咽拭子标本，评价方法的准确性。 结果：LAMP技术最佳反应温度为65 ℃，临床分离的40株SA检测阳性率达100%；其他菌株均无非特异性反应；最低检测限14 CFU/test。以培养法为标准， LAMP试验检测60份咽拭子标本的敏感性、特异性、阳性预测值和阴性预测值为87.5%、100%、100%和98.1%。 结论：LAMP技术操作简便、快速、灵敏度高、特异性强，可于60 min内快速筛查咽拭子标本中SA。
Abstract: Objective：To establish a method of loopmediated isothermal amplification assay (LAMP) for rapid detecting Staphylococcus aureus (S.aureus) in clinical samples. Methods：A highly specific set of four primers, two inner primers (the forward inner primer termed FIP and the backward inner primer termed BIP) and two outer primers (F3 and B3) that recognize a total of six distinct sequences at target DNA, was designed to detect femA gene ofS.aureus and LAMP forS.aureus was developed. Forty clinical isolates of S.aureus, 12 other grampositive cocci and 8 kinds of gramnegative bacteria was detected, and the specificity of the method was evaluated. The standard stain ofS.aureus ATCC 25923 was diluted by 10n, and the sensitivity was evaluated. S.aureus in 60 clinical throat swab samples was detected by LAMP and culture method was performed respectively. Results：The optimal incubation temperature of LAMP was 65 ℃. The 40 clinical isolates of S.aureus were detected by LAMP and the positive rate was 100%. No specific reactions were found in other bacteria stains. The LAMP assay showed a limit of 14 CFU/test. The sensitivity，specificity，positive predictive value and negative predictive value of LAMP by detecting sixty throat swab samples was 87.5%，100%，100% and 98.1%, respectively. Conclusion：The developed LAMP is very simple, rapid, sensitive and specific method and is available for rapid identification for clinic swab specimen within 60 min.