应用基因芯片检测系统诊断结核分枝杆菌感染及耐药基因突变的临床研究

Clinical application of genechip based detection system in diagnosing mycobacterium infection and TB drug resistance gene mutation

目的探讨基因芯片检测系统在分枝杆菌属鉴定及耐药性分析的临床应用价值。方法应用基因芯片技术,对患者的痰液、穿刺脓液、胸腹水、脑脊液等标本进行分枝杆菌属DNA检测,并进一步对结核分枝杆菌(MTB)阳性标本进行耐药性分析。同时采用抗酸染色检测标本,比较两种方法检出率。采用MTB感染T细胞IFN-γ释放试验(TB-IGRA)技术对部分患者进行结核感染检测。结果 4 402例患者标本中检出分枝杆菌148例,检出率为3.36%(148/4 402)。其中MTB复合群137例,非结核分枝杆菌(NTM)11例,NTM占分枝杆菌总检出例数的7.4%(11/148)。检出率较高的标本为穿刺脓液、肺泡灌洗液和组织。进一步对137例MTB阳性标本进行耐利福平基因rpoB及耐异烟肼katG、inhA基因检测,共有22例检测到了突变的耐药基因;基因芯片法检测痰液、脑脊液和胸腹水分枝杆菌检出率明显高于抗酸染色法(P<0.05)。基因芯片检测结核感染的检出率明显低于TB-IGRA(P<0.05)。结论基因芯片检测系统可以直接进行分枝杆菌属鉴定及耐药性分析,尤其适用于痰液、胸腹水和脑脊液。该方法简便快速,灵敏度较高,特异性好。

Objective To investigate the clinical application value of genechip detection system in the mycobacterial species identification and drug resistance analysis.Methods The specimens of sputum,punctured pus,pleural and abdominal ascites,cerebrospinal fluid and so on were performed the Mycobacterium DNA detection by using the gene chip technique.Then Mycobacterium tuberculosis positive samples were further performed the drug-resistant analysis.Meanwhile the Ziehl-Neelsen acid-fast staining was adopted to detect the sample.The positive rates were compared between the two groups.And TB-IGRA was used to examine the tubercle bacillus infection in partial patients.Results In 4 402 samples,137cases（3.36%）of Mycobacterium tuberculosis（MTB）and 11cases（7.4%）of non-tuberculosis mycobacterium（NTM）were detected.Puncture solution,bronchoalveolar lavage fluid and tissue specimen had higher positive rate.In the 137 positive MTB samples by rifampicin-resistant gene rpoB,isoniazide-resistant gene katG and inhA detection,22 cases of resistance gene mutations were detected;the positive rate of genechip for detecting sputum,cerebrospinal fluid,hydrothorax and ascites was higher than that of acid-fast staining.TB-IGRA detection had higher positive rate of TB infection than genechip.Conclusion The genechip detection system can directly conduct Mycobacterium identification and drug resistance analysis,which is especially suitable for sputum,cerebrospinal fluid,hydrothorax and ascites samples,and which is simple and rapid with higher sensitivity and good specificity.