新型胶颗粒芯片技术检测痰标本中结核分枝杆菌及其耐药性的研究

New TruArray technology for detection of Mycobacterium tuberculosis and drug resistance from sputum specimen

目的评估一种新型胶颗粒芯片技术[TruArray胶颗粒芯片（Gel element arrays），简称为“TruArray芯片”]用于痰标本中结核分枝杆菌及利福平（RFP）和异烟肼（INH）耐药性的检测效能。方法选取2016年8月在首都医科大学附属北京胸科医院就诊的149例疑似肺结核患者痰标本，分别进行涂片镜检、BACTEC MGIT 960快速液体培养系统（简称“MGIT 960”）、TruArray芯片和GeneXpert MTB／RIF（简称“GeneXpert”）检测，对液体培养阳性菌株进行测序菌种鉴定、MGIT960药物敏感性试验（简称“药敏试验”）和耐药相关基因测序，分析TruArray芯片检测痰标本中结核分枝杆菌及其耐药性的检测效能。结果液体培养、GeneXpert和TruArray芯片对结核分枝杆菌的检出率分别为32．2％（48／149），53．7％（80／149）和57．0％（85／149），TruArray芯片和GeneXpert两种方法检出率差异无统计学意义（χ2-0．34，P-0．560），但TruArray芯片检测阳性率高于液体培养（χ^2=18．59，P〈0．01）。以MGIT960试验结果为标准，TruArray芯片检测痰标本中结核分枝杆菌的敏感度和特异度分别为97．90％（47／48）和61．4％（62／101）；以GeneXpert检测结果为标准，TruArray芯片检测结核分枝杆菌的敏感度和特异度分别为97．5％（78／80）和88．4％（61／69）。以MGIT960药敏试验结果为标准，TruArray芯片检测RFP耐药的敏感度和特异度分别为91．7％（11／12）和96．8％（30／31）；以GeneXpert检测结果为标准，TruArray芯片检测RFP耐药的敏感度和特异度为85．0％（17／20）和97．9％（47／48）；以耐药基因测序结果为标准，TruArray芯片检测RFP耐药的敏感度和特异度分别为92．3％（12／13）和100．0％（30／30）。以MGIT960液体药敏试验结果为标准，TruArray芯片技术检测INH耐药的敏感度和特异度分别为78．6％（11／14）和93．8％（30／32）；以耐药基因测序结果为判断标准，TruArray芯片检测INH耐药的敏感度和特异度分别为100．0％（11／11）和94．3％（33／35）。结论TruArray芯片检测技术快速、可靠，在结核病及耐多药结核病快速诊断方面具有非常好的应用前景。

Objective To evaluate the performance of a new gel element arrays gel-drop （TruArray） chip for the detection of Mycobacterium tuberculosis （MTB） and rifampicin （RFP） and isoniazid （INH） resistance from sputum specimen. Methods A total of 149 samples provided by pulmonary tuberculosis suspected patients from Capital Medical University Affiliated Beijing Chest Hospital were selected for smear microscopy, MGIT 960 liquid culture, TruArray and GeneXpert MTB/RIF （GeneXpert） test, and perform identification sequencing, MGIT 960 drug susceptibility test and drug resistance-related gene sequencing for liquid culture positive strains. Analyze the performance of TruArray test for MTB and its drug resistance detection in sputum samples. Results The detec tion rates of MTB in liquid culture, GeneXpert and TruArray test were 32.2% （48/149）, 53.7% （80/149） and 57.0% （85/149）, respectively. There was no statistical difference in the detection rates of TruArray and GeneXpert （χ2 =0.34, P=0. 560）, but the positive rate of TruArray test was significantly higher than that of liquid culture（χ2= 18.59, P=0.01）. With the MGIT 960 liquid culture result as standard, the sensitivity and specificity of TruArray test for MTB detection in sputum samples were 97.9% （47/48） and 61.4% （62/101）, respectively. Compared with GeneXpert result, the sensitivity and specificity of TruArray test for MTB detection were 97.5G （78/80） and 88. 4% （61/69）, respectively. Based on the results of MGIT 960 drug susceptibility test, the sensitivity and specificity of TruArray test for RFP resistance detection were 91.70//oo （11/12） and 96.8G （30/31）, respectively. Compared with GeneXpert assay, the sensitivity and specificity of the TruArray test for RFP resistance detection were 85.0G （17/20） and 97.9-/00 （47/48）, respectively. Based on the sequencing results of drug resistance genes, the sensitivity and specificity of TruArray test for RFP resistance detection was 92.3% （12/13） and 100. 0% （30/30）, respectively. Based on the results of MGIT 960 liquid susceptibility test, the sensitivity and specificity of the TruArray test for INH resistance detection were 78.6% （11/14） and 93.8G （30/32）, respectively. Using the sequencing results of drug resistance genes as the criterion, the sensitivity and specificity of TruArray test for INH resistance detection were 100.0% （11/11） and 94.3% （33/35）, respectively.