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目的 评价二代测序技术在活动性结核病患者中的诊断价值.方法 前瞻性纳入2017年3月1日至2017年9月1日来华山医院就诊的临床怀疑活动性结核病的患者34例,在华山医院进行二代测序,并跟踪随访临床诊疗结局.送检标本(支气管灌洗液4例,脑脊液18例,胸腔积液4例,脓液3例,腹腔积液、肝组织各2例,关节积液1例)均在BGISEQ-100平台上行二代测序,测序所得病原序列和病原数据库进行比对得到最终结果.二代测序结果以检测到结核分枝杆菌复合群唯一匹配序列为阳性,未检测到唯一匹配序列为阴性.以符合各类标本结核分枝杆菌培养阳性、病理诊断为结核感染、Xpert MTB/RIF检测阳性及结核分枝杆菌核酸检测阳性等4项中至少1项即为确诊活动性结核病患者;临床可疑活动性结核病且抗结核药物治疗有效为临床诊断患者;有其他病原学依据或临床排除活动性结核病者为非结核感染.分析二代测序在活动性结核病诊断中的敏感度、特异度.结果 确诊为活动性结核病患者9例,其中Xpert MTB/RIF检测阳性6例,病理诊断为结核感染4例,培养阳性3例,结核核酸PCR检测阳性1例;临床诊断为活动性结核病患者10例,非结核感染患者15例.在确诊及临床诊断患者中,二代测序技术检测到结核分枝杆菌复合群序列11例,敏感度为57.9％(11/19),特异度为100.0％(15/15).在确诊患者中,二代测序技术的敏感度为66.7％(6/9);在同步进行Xpert MTB/RIF检测与二代测序的31例标本中,以临床诊断为金标准,两者的特异度均为100.0％(15/15),敏感度分别为37.5％(6/16)与50.0％(8/16),两者比较差异无统计学意义(McNemar test,P=0.727).结论 二代测序技术能够较为快速地检测多类标本中的结核分枝杆菌复合群,且其敏感度及特异度与Xpert MTB/RIF相当,可以作为确诊试验用于活动性结核病的早期辅助诊断.“,”Objective To evaluate the diagnostic value of next-generation sequencing (NGS) in active tuberculosis infection.Methods The clinical suspicion of active tuberculosis with sample submitted for NGS platform in Huashan Hospital was prospectively enrolled in the period from March 1,2017 to September 1,2017.All the cases were followed up for final clinical diagnosis.The submission samples (4 cases of bronchial lavage fluid,18 cases of cerebrospinal fluid,pleural effusion in 4 cases,3 cases of pus,ascites,liver tissue in 2 cases,1 case of synovial fluid) were all sequenced on the BGISEQ-100 platform,and the final result was obtained by comparing the pathogenic sequences to the pathogen database.A positive result for NGS was defined as detection of the single matching sequence of Mycobacterium tuberculosis complex At least one of tuberculosis culture,pathologically diagnosed tuberculosis infection,Xpert MTB/RIF positive and tuberculosis nucleic acid test is positive for the diagnosis of definite active tuberculosis infection.There are other etiological evidence or clinical exclusion of active tuberculosis infection as non-tuberculosis infection.Then we analyze the sensitivity and specificity of NGS in the diagnosis of tuberculosis.Results Among them,9 cases was definite TB cases,including Xpert MTB/RIF positive in 6 cases,pathological diagnosis in 4 cases,3 cases of culture positive,tuberculosis nucleic acid PCR positive in 1 case;clinical diagnosis of active tuberculosis in 10 cases,non-tuberculosis infection in 15 cases.Among the definite and clinically diagnosed 1cases,11 cases of Mycobacterium tuberculosis complex were detected by NGS.The sensitivity and specificity were 57.9％ (11/19) and 100.0％ (15/15) respectively.In confirmed cases,the NGS sensitivity was 66.7％ (6/9).Among the 31 specimens sent in parallel for Xpert MTB/RIF and NGS,the clinical diagnosis was the gold standard,with a specificity of 100.0％ (15/15) and a sensitivity of 37.5％ (6/16) and 50.0％ (8/16),respectively,with no significant difference (McNemar test,P=0.727).Conclusion The second-generation sequencing can detect Mycobacterium tuberculosis complex in multiple samples more rapidly.Its sensitivity and specificity are comparable to those of Xpert MTB/RIF and can be used as a diagnostic test for the early diagnosis of active TB.