利用基因芯片检测结核分支杆菌及其利福平耐药性的研究

目的 探讨结核杆菌耐利福平(RFP)检测基因芯片在结核病诊断及其耐药性检测中的应用价值。方法 采用结核杆菌耐RFP芯片对35株RFP耐药的临床分离菌株，102例肺结核患者、27例非结核病的其他患者的痰标本中的结核杆菌及其RFP耐药性进行了检测，基因芯片检测结果与痰涂片、细菌培养及结核杆菌标准化药敏试验结果比较。结果结核杆菌耐RFP检测芯片检测35株耐药株，有33株判定为RFP耐药株，与传统药敏试验结果的符合率为94．29％。对27份其他患者的痰液标本进行结核杆菌检测，特异性为92．59％。对102份结核患者痰标本中结核杆菌进行检测，痰涂片法阳性率35．29％(36／102)，细菌培养法阳性率28．43％(29／102)，基因芯片法阳性率77．45％(79／102)。传统的药敏试验报告102份痰标本仅29份培养出结核分支杆菌，其中8份RFP耐药株，而基因芯片法检测102份痰标本中发现20份耐RFP结核杆菌。主要基因突变位点为531、526和516。结论 采用结核杆菌耐RFP检测芯片检测结核杆菌及RFP耐药性具有快速简便、特异性高的特点。

Identification of Mycobacterium tuberculosis and rifampin-resistant strains by gene-chips

Objective To evaluate the gene-chip detecting rifaman-resistance Mycobacterium tuberculosis applied in TB diagnosis and drug-resistant dectection. Methods Mycobacterium tuberculosis and rifaman-resistant strains among 35 rifaman-resistance isolated strains and 102 sputa specimens from TB patients,27 sputa specimens from other patients were examined the gene-chips. Results obtained were compared with sputum examination, bacteriological culture and standard drug susceptibility test of Mycobacterium tuberculosis . Results Thirty-five rifaman-resistance strains were detected by gene-chips and 33 were identified as rifaman-resistance strains and the concordance with the traditional drug susceptibility test of Mycobacterium tuberculosis was 94.29 %. Twenty-seven sputa specimens from other patients were examined Mycobacterium tuberculosis by the gene-chips, 2 were positive, the detection specialty was 92.59 %. Using three methods detecting Mycobacterium tuberculosis among 102 sputa specimens the positive rate respectively was, sputum examination 35.29 % (36/102), bacteriological culture 28.43 % (29/102), gene-chip 77.45 % (79/102). Among 102 sputa specimens only 29 examined Mycobacterium tuberculosis by the traditional drug susceptibility test and 8 were rifaman-resistant strains. While using gene-chip, there were 20 among 102 sputa specimens identified as rifaman-resistance strains. Among total 55 rifaman-resistance strains detected by the gene-chips, the most frequent mutations were those associated with codon 531 (23 of 55; 41.8 %), 526(15 of 55; 27.27 %) and 516(9 of 55; 16.36 %). Conclusion Results showed that this was a rapid, simple and highly specific method when using gene-chip to detect Mycobacterium tuberculosis and rifaman-resistant strains.