Cheng Jie, Zheng Song, Peng Ling, Li Mei, Wang Dianchao, Li Yong, Ma Rong
Clin Lab. 2024 Dec 1;70(12). doi: 10.7754/Clin.Lab.2024.240532.
Early diagnosis of Mycobacterium tuberculosis (MTB) infection is of great significance for the clinical management of tuberculosis (TB). We first explored the efficacy of single-molecule nanopore DNA sequencing in the early diagnosis of suspected TB patients and analyzed the advantages in differentiating and diagnosing MTB and non-tuberculous Mycobacteria (NTM).
In this cohort study, we reviewed the clinical data of suspected TB patients admitted from December 1, 2021, through April 15, 2022. All patients underwent 3 - 6 times acid-fast bacilli smear examinations of sputum, all of which were negative. To make a definitive diagnosis, we extracted specimens from the patients and performed specimen culture, Xpert MTB/Rif assay, and single-molecule nanopore DNA sequencing. The efficacy of different diagnostic methods in diagnosing suspected TB patients was compared using "Diagnostic Criteria for Pulmonary Tuberculosis" (WS288-2017) as the gold standard.
Among the 25 patients, 15 were infected with MTB, 5 were infected with NTM, 1 had mixed MTB and NTM infection, and 4 were negative. The accuracy of single-molecule nanopore DNA sequencing in diagnosing mycobacterial infection (MTB + NTM) was 92.0%, with a sensitivity of 90.5% and a specificity of 100%; the accuracy of diagnosing MTB infection was also 92.0%, with a sensitivity of 87.5% and a specificity of 100%. Single-molecule nanopore DNA sequencing showed an accuracy of 100% in differentiating MTB and NTM. However, the diagnostic accuracy and sensitivity of specimen culture and Xpert MTB/Rif assay were relatively low (≤ 52%) compared to "specimen culture + Xpert MTB/Rif assay". The diagnostic efficacy of single-molecule nanopore DNA sequencing was not affected by the source of tissue samples, while specimen culture and Xpert MTB/Rif assay could not diagnose mycobacterial infection using extrapulmonary specimens.
As a third-generation sequencing technology, single-molecule nanopore DNA sequencing has significant application value in diagnosing suspected TB patients. Compared to traditional diagnostic methods, such as specimen culture and Xpert MTB/Rif assay, single-molecule nanopore DNA sequencing exhibits high diagnostic efficacy, low error rate, and convenient detection.
结核分枝杆菌（MTB）感染的早期诊断对结核病（TB）的临床管理具有重要意义。我们首次探讨了单分子纳米孔DNA测序在疑似结核病患者早期诊断中的效果，并分析了其在鉴别诊断MTB和非结核分枝杆菌（NTM）方面的优势。
在这项队列研究中，我们回顾了2021年12月1日至2022年4月15日收治的疑似结核病患者的临床资料。所有患者均接受了3 - 6次痰涂片抗酸杆菌检查，结果均为阴性。为明确诊断，我们从患者身上提取标本并进行标本培养、Xpert MTB/Rif检测和单分子纳米孔DNA测序。以《肺结核诊断标准》（WS288 - 2017）作为金标准，比较不同诊断方法对疑似结核病患者的诊断效果。
25例患者中，15例感染MTB，5例感染NTM，1例为MTB和NTM混合感染，4例为阴性。单分子纳米孔DNA测序诊断分枝杆菌感染（MTB + NTM）的准确率为92.0%，灵敏度为90.5%，特异度为100%；诊断MTB感染的准确率也为92.0%，灵敏度为87.5%，特异度为100%。单分子纳米孔DNA测序在鉴别MTB和NTM方面的准确率为100%。然而，与“标本培养 + Xpert MTB/Rif检测”相比，标本培养和Xpert MTB/Rif检测的诊断准确率和灵敏度相对较低（≤52%）。单分子纳米孔DNA测序的诊断效果不受组织样本来源的影响，而标本培养和Xpert MTB/Rif检测无法使用肺外标本诊断分枝杆菌感染。
作为第三代测序技术，单分子纳米孔DNA测序在诊断疑似结核病患者方面具有重要应用价值。与标本培养和Xpert MTB/Rif检测等传统诊断方法相比，单分子纳米孔DNA测序具有诊断效率高、错误率低、检测方便等优点。
