皮肤结核快速诊断中环介导等温扩增技术的建立  

Rapid Diagnosis of Cutaneous Tuberculosis with Loop-mediated Isothermal Amplification

作　　者：冯雨苗[1] 王淼淼[2] 王洪生 FENG Yumiao;WANG Miaomiao;WANG Hongsheng(Department of Dermatology,the Affiliated Suzhou Hospital of Nanjing Medical University,Suzhou 215001,China;Department of Dermatology,the First Affiliated Hospital of Soochow University,Suzhou 215006,China;Laboratory of Leprosy and Other Mycobacterial Infections,Institute of Dermatology,Chinese Academy of Medical Sciences,Nanjing 210042,China)

机构地区：[1]南京医科大学附属苏州医院皮肤科,江苏苏州215001 [2]苏州大学第一附属医院皮肤科,江苏苏州215006 [3]中国医学科学院皮肤病研究所麻风分枝杆菌实验室,江苏南京210042

出　　处：《中国皮肤性病学杂志》2019年第9期1097-1100,共4页The Chinese Journal of Dermatovenereology

摘　　要：目的建立一种用于皮肤结核标本快速检测的环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)。方法根据结核分枝杆菌复合群gyrB基因序列设计三对特征性引物,采用LAMP对结核分枝杆菌进行65 ℃等温扩增。评价该方法的灵敏度和特异性,并初步探讨其在临床皮肤结核标本中的应用。结果建立的LAMP检测结核分枝杆菌的灵敏度与PCR相当;与其他分枝杆菌无交叉反应,特异性良好。针对培养阳性的皮肤结核标本,LAMP检出3例(3/5),略优于PCR(2/5),且LAMP电泳条带更清晰易辨认。结论LAMP技术操作简单,敏感性和特异性较强,可应用于皮肤结核标本的快速检测。Objective To develop loop-mediated isothermal amplification for rapid detection of cutaneous tuberculosis specimens.Methods The specific primers were designed by targeting the gyrB gene of M.tuberculosis.LAMP was performed at 65 ℃.The sensitivity and specificity of LAMP was evaluated.Results The sensitivity of LAMP was comparable to that of PCR.There was no crossing-reaction with other mycobacterium.In five clinical culture-positive samples,the detection rate of LAMP was 60%,slightly higher than PCR (40%),and electrophoretic bands of LAMP were easier to identify.Conclusion LAMP is simple,sensitive,specific and has potential for rapid diagnosis of cutaneous tuberculosis.

关 键 词：环介导等温扩增技术 结核分枝杆菌 聚合酶链反应 皮肤结核 

分 类 号：R529.4[医药卫生—内科学]