RT-LAMP与LAMP法检测结核分枝杆菌的效果比较  

Comparision on the effect of RT-LAMP and LAMP method for detection of Mycobacterium tuberculosis

作　　者：吴丹丹 李保胜[2] 亢继文[2] 孙殿兴[2] WU Dan-dan;LI Bao-sheng;KANG Ji-wen;SUN Dian-xing(Chengde Medical University,Chengde 067000,China;Liver and Infectious Diseases Department,Bethune International Peace Hospital of PLA, Shijiazhuang 050082,China)

机构地区：[1]承德医学院,承德067000 [2]中国人民解放军白求恩国际和平医院肝病传染科,石家庄050082

出　　处：《中国人兽共患病学报》2018年第3期248-254,共7页Chinese Journal of Zoonoses

基　　金：全军医药卫生科研基金项目(No.CBJ14C010)~~

摘　　要：目的比较RT-LAMP、LAMP、L-J法检测MTB的效果,为结核病的快速诊断提供依据。方法用常见的非结核分枝杆菌与其他常见的呼吸道感染病菌(金黄色葡萄球菌、铜绿假单胞菌、肺炎克雷伯杆菌)检测特异性,并用酶切实验对结核分枝杆菌特异性产物进行酶切鉴定;用RT-LAMP、LAMP、L-J法检测100名结核病患者和22名来自无结核病患者痰标本中的结核分枝杆菌测试灵敏性;对浓度为1ng/μL H37Rv标准株进行对倍比稀释用于RT-LAMP极限检测实验。结果RT-LAMP、LAMP、L-J方法的阳性率分别为100%,92%和88%;RT-LAMP和L-J,RT-LAMP和LAMP差异均有统计学意义;RT-LAMP的灵敏度比LAMP高10倍;RT-LAMP不仅可以识别活菌,并且能够重复检测MTB的单一拷贝。结论 RTLAMP检测效果优于LAMP及L-J,适于基层医院结核病诊断的推广和应用。We compared the RT-LAMP,LAMP and L-J for detecting MTB to provide the rapid diagnosis method for tuberculosis.In this assay,NTM and other common respiratory bacteria were used to detect specificity,Mycobacterium tuberculosis specific products were identified by restriction enzyme digestion.To detect sensitivity,we used RT-LAMP,LAMP and L-J to detect 100 cases sputum specimens from patients with tuberculosis and 22 cases control sputum specimens,the 10 times dilution of 1 ng/μL H37Rv standard strains was used to detect RT-LAMP limit.The results showed that the positive rate of RT-LAMP,LAMP and L-J were 100%,92%,88%.RT-LAMP and L-J,RT-LAMP and LAMP were statistically significant,RT-LAMP had 10 times higher sensitivity than LAMP,RT-LAMP not only to identify viable but also capable of detecting a single copy of MTB.So RT-LAMP is superior to LAMP and L-J and is practical for use in primary medical care institution or peripheral laboratory.

关 键 词：结核分枝杆菌 逆转录-环介导等温扩增(RT-LAMP) 环介导等温技术(LAMP) 16S RRNA 

分 类 号：R378.91[医药卫生—病原生物学]