肺外结核感染的快速分子检测和菌株分型方法的应用  

Rapid molecular assay and strain typing methods of extrapulmonary tuberculosis

作　　者：郭建[1] 范齐文[1] 卢水华[2] 邓桂林[2] 吴文娟[1] 

机构地区：[1]同济大学附属东方医院南院医学检验科,上海200120 [2]上海市公共卫生临床中心医学检验科

出　　处：《中华检验医学杂志》2013年第7期620-624,共5页Chinese Journal of Laboratory Medicine

基　　金：上海市科委引导类课题资助项目（114119a0200）

摘　　要：目的应用结核分枝杆菌rRNA直接检测（MTD）和多位点PCR菌株分型方法，检测肺外结核标本中的结核分枝杆菌，评价其在肺外结核感染的快速检测和菌株分型中的应用价值。方法回顾性分析2010年6月至2011年12月上海市公共卫生临床中心47例儿童肺外结核、75例成人肺外结核以及48例非结核病患者的脑脊髓液、穿刺液等标本。用涂片法、L-J固体培养法、MGIT960液体培养法和MTD法进行平行检测，对培养阳性菌株应用MPT64胶体金初筛。MPT64阴性菌株采用多位点PCR菌株分型方法进行快速分型，并用化学法确认。采用SPSSl6．0进行统计分析。结果涂片法、L-J固体培养法、MGIT960液体培养法和MTD法检测肺外结核的敏感度依次为：10．7％（13／122），11．5％（14／122），16．4％（20／122）和37．7％（46／122），特异度均为100．0％。20株培养阳性的菌株，其中6例儿童患者临床分离菌株为MPT64胶体金检测阴性，经多位点PCR菌株分型方法、化学法分型鉴定为卡介苗菌株。多位点PCR菌株分型方法与传统化学法结果一致，但可在4h内完成检测。结论与传统病原学检测方法比较，MTD法检测结核分枝杆菌快速简便，结果灵敏可靠；同时应用多位点PCR菌株分型方法，可对儿童肺外结核患者进行快速的早期鉴别诊断。Objective To evaluate the effect of Mycobacterium tuberculosis Direct Assay （MTD） for rapid detecting Mycobacterium tuberculosis rRNA and Multi-locus PCR for M. boris BCG strain typing in patients with suspected extra-pulmonary tuberculosis. Methods From June 2010 to December 2011, 47 children and 75 adult patients with suspected extra-pulmonary tuberculosis in Shanghai public health clinical center were recruited. Also 48 non-tuberculosis patients were taken as a negative control. Clinical specimens from these patients were collected. Acid fast stain, solid culture, liquid culture, and MTD were used to detect all clinical specimens simultaneously. Screen tuberculosis strains of the culture isolates by MPT64 antigen assay and use Multi-locus PCR for the BCG strain genotyping of the isolates without MPT64 antigen. SPSS16.0 was used to analyse the results. Results The sensitivity for acid fast stain, solid culture, liquid culture and MTD test was 10. 7% （ 13/122）, 11.5% （ 14/122）, 16. 4% （20/122） and 37.7% （46/ 122）, respectively. And the specificity of MTD was 100. 0%. Six clinical isolates from children were identified as BCG by Muhi-locus PCR typing, the same with chemical tests. Conclusions The MTD assay and the MGIT960 liquid culture are effective and reliable method for diagnosing extra-pulmonac~ tuberculosis. And Multi-locus PCR can be assisted for the early diagnosis of extra-pulmonary tuberculosis patients with suspected BCG infection.

关 键 词：分枝杆菌 结核 RNA 核糖体 细菌分型技术 聚合酶链反应 

分 类 号：R512.5[医药卫生—内科学]