Objectives: To assess the value of next-generation sequencing (NGS) technology in early diagnosis of patients with tuberculous meningitis (TBM).

Methods: 56 patients with clinically suspected TBM who came to Shandong Provincial Chest Hospital from February 2, 2018 to August 2, 2018 were prospectively included, and the clinical diagnosis and treatment outcomes were followed up. NGS was performed for the cerebrospinal fluid specimens submitted for test on the BGISEQ-100 platform of Tianjin Huada Gene Research Institute and the obtained pathogen sequences were compared with the pathogen data to get the final results. The NGS results were positive for detecting the unique matching sequence of the Mycobacterium tuberculosis (MTB) complex and negative for no unique matching sequence. Patients confirmed with TBM should have at least one of the following four items: cerebrospinal fluid MTB culture positive, smear positive, Xpert MTB/RIF test positive, or MTB nucleic acid polymerase chain reaction (PCR) test positive; clinically diagnosed patients were those with clinically suspected TBM and effective anti-tuberculosis treatment; non-TBM patients were those with other pathogenic basis or clinical exclusion of TBM. The sensitivity and specificity of NGS in early diagnosis of TBM were analyzed.

Results: 22 patients were confirmed with TBM, of which 13 were positive for Xpert MTB/RIF test, 6 were positive for cerebrospinal fluid MTB culture, 5 were positive for MTB nucleic acid PCR test, 12 patients were clinically diagnosed with TBM, and there were 16 cases of non-TBM patients. Among confirmed and clinically diagnosed patients, 20 cases of MTB complex were detected by NGS technology, with a sensitivity of 58.8% (20 / 34) and specificity of 100% (16 / 16). Among confirmed patients, the sensitivity of NGS was 63.6% (14 / 22). Of the 50 specimens that were simultaneously subjected to traditional methods, Xpert MTB/RIF test and NGS, the specificity of the three methods was 100% (16 / 16) based on clinical diagnosis, and the sensitivity was 29.4% (10 / 34), 38.2% (13 / 34), and 58.8% (20 / 34) respectively. The difference of sensitivity between the first two detection methods and NGS was statistically significant (McNemar test, p = 0.013, x² = 5.786 and p = 0.065, x² = 3.273). The sensitivity of traditional methods combined with NGS was as high as 82.4% (28 / 34).

Conclusions: NGS technology could rapidly detect the MTB complex in cerebrospinal fluid with significant sensitivity and specificity, which could be used as an early diagnosis index of TBM. NGS combined with MTB culture could increase the detection rate.

目的：评估下一代测序（NGS）技术在结核性脑膜炎（TBM）患者早期诊断中的价值。

方法：前瞻性纳入2018年2月2日至2018年8月2日在山东省胸科医院就诊的56例临床疑似TBM患者，并对其临床诊断和治疗结果进行随访。在天津华大基因研究所的BGISEQ-100平台上对提交测试的脑脊液标本进行NGS，并将获得的病原体序列与病原体数据进行比较，以获得最终结果。NGS结果对于检测结核分枝杆菌的独特匹配序列是阳性的（MTB）复杂且为负数，表示没有唯一的匹配序列。确认患有TBM的患者应至少具有以下四项中的一项：脑脊液MTB培养阳性，涂片阳性，Xpert MTB / RIF测试阳性或MTB核酸聚合酶链反应（PCR）测试阳性；临床确诊的患者是那些临床上疑似TBM且有效抗结核治疗的患者；非TBM患者是具有其他病因基础或临床排除TBM的患者。分析了NGS在TBM早期诊断中的敏感性和特异性。

结果：确诊为TBM的22例患者，其中Xpert MTB / RIF检测阳性13例，脑脊液MTB培养阳性6例，MTB核酸PCR检测阳性5例，临床诊断为TBM的12例，其中16例非TBM患者的病例。在已确诊和临床诊断的患者中，通过NGS技术检测到20例MTB复合体，敏感性为58.8％（20/34），特异性为100％（16/16）。在确诊的患者中，NGS的敏感性为63.6％（14/22）。在Xpert MTB / RIF测试和NGS同时接受传统方法的50个标本中，根据临床诊断，这三种方法的特异性为100％（16/16），敏感性为29.4％（10/34） ，38.2％（13/34）和58.8％（20/34）。p  = 0.013，x ²  = 5.786，p  = 0.065，x ²  = 3.273）。传统方法与NGS结合使用的灵敏度高达82.4％（28/34）。

结论： NGS技术可以快速，快速，准确地检测出脑脊液中的MTB复合物，可作为TBM的早期诊断指标。NGS与MTB培养相结合可以提高检出率。