To evaluate the early diagnostic value of next-generation sequencing technology applied to cerebrospinal fluid detection in patients with tuberculous meningitis.

A total of 50 patients with clinically suspected tuberculous meningitis who were treated in Shandong Provincial Chest Hospital from February 2nd, 2018 to August 2nd, 2018 were collected prospectively, and the diagnosis and treatment outcomes of those patients were followed up. The submitted cerebrospinal fluid specimens were all subjected to next-generation sequencing, and the obtained original sequence was compared with the pathogenic microorganism database to get the final results. The next-generation sequencing results showed positive when detecting the unique alignment sequence of the Mycobacterium tuberculosis complex, and negative when no unique alignment sequence was detected. Tuberculous meningitis were confirmed with at least one of the four items: cerebrospinal fluid Mycobacterium tuberculosis culture positive, smear positive, Xpert MTB/RIF test positive and Mycobacterium tuberculosis nucleic acid test positive; clinically diagnosed tuberculous meningitis patients were with clinically suspected tuberculous meningitis and effective anti-tuberculosis treatment; non-tuberculous meningitis patients were with other etiological evidence or clinical exclusion of tuberculous meningitis. The sensitivity and specificity of next-generation sequencing in early diagnosis of tuberculous meningitis were analyzed, respectively.

Among the 22 patients with confirmed tuberculous meningitis, 13 cases were positive for Xpert MTB/RIF test, 6 cases were positive for culture, and 5 cases were positive for Mycobacterium tuberculosis nucleic acid by PCR. There were 12 cases clinically diagnosed as tuberculous meningitis and 16 cases as non-tuberculous meningitis patients. Among the confirmed and clinically diagnosed patients, 20 cases of Mycobacterium tuberculosis complex series were detected by next-generation sequencing technology, with a sensitivity of 58.8% (20/34) and a specificity of 100% (16/16). Among the confirmed patients, the sensitivity of next-generation sequencing was 63.6% (14/22). Among the 50 specimens that were simultaneously submitted for Mycobacterium tuberculosis culture, Xpert MTB/RIF test and next-generation sequencing, the specificity of the three methods was 100% (16/16) with clinical diagnosis as the standard. The sensitivity of traditional method, Xpert MTB/RIF test and next-generation sequencing were 29.4% (10/34), 38.2 (13/24) and 58.8 (20/34), respectively. The sensitivity differences between the first two detection methods and next-generation sequencing were significantly different (McNemar test: χ² = 8.333, P = 0.013; χ² = 8.333, P = 0.065). The sensitivity of the combined detection of traditional method and next-generation sequencing was as high as 82.4% (28/34).

Next-generation sequencing technology could quickly detect the Mycobacterium tuberculosis complex in the cerebrospinal fluid, with significant sensitivity and specificity, and could be used as an early diagnosis index for tuberculous meningitis. Next-generation sequencing combined with traditional detection method could increase the detection rate.