Diagnosing tuberculous meningitis from cell-free DNA by multi-targeted real-time PCR: An experience from 170 cases

IF 2.9 3区医学 Q3 IMMUNOLOGY

Tuberculosis Pub Date : 2025-12-01 Epub Date: 2025-10-29 DOI:10.1016/j.tube.2025.102705

Kusum Sharma , Megha Sharma , Ritu Shree , Neeraj Singla , Himanshu Joshi , Ashish Kumar Kakkar , Apinderpreet Singh , Aman Sharma , Navneet Sharma , Manish Modi

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引用次数: 0

Abstract

Setting

and objective: To evaluate the diagnostic utility of Cell-free DNA (Cf-DNA)-based multi-targeted real-time PCR (Mrt-PCR) for diagnosing tuberculous meningitis, the most severe form of extrapulmonary tuberculosis.

Design

A total of 170 cerebrospinal fluid samples (CSF) of tuberculous meningitis [Definite TBM (n = 146), Probable TBM (n = 24)] and 100 non-TB controls were subjected to Cf-DNA Mrt-PCR test using IS6110, IS1081, and nrdZ gene targets. The performance was also evaluated against Truenat MTB Plus assay and GeneXpert MTB/RIF Ultra.

Results

The sensitivity, specificity, positive predictive value and negative predictive value of Cf-DNA M-rtPCR was 78.24 %, 100 %, 100 % and 72.99 %, respectively in overall diagnosis of TBM and 82.88 %, 100 %, 100 % and 80 %, respectively in diagnosing ‘definite’ TBM. Cf-DNA M-rtPCR detected 14 additional cases that were missed by Truenat MTB Plus assay and GeneXpert MTB/RIF Ultra. Among the different gene targets used, eight cases were detected only by IS1081, three only by IS6110 and one only by nrdZ gene. Six cases were reported rifampicin-resistant by both Truenat MTB Plus assay and GeneXpert MTB/RIF Ultra and were also confirmed by rpoB sequencing.

Conclusion

The detection of cell-free DNA, along with M-rtPCR could serve as a reliable tool for diagnosing tuberculous meningitis directly from CSF samples.

查看原文本刊更多论文

多靶点实时PCR检测游离DNA诊断结核性脑膜炎170例的经验。

背景和目的：评估基于无细胞DNA （Cf-DNA）的多目标实时PCR （Mrt-PCR）诊断结核性脑膜炎（最严重的肺外结核形式）的诊断效用。设计：采用IS6110、IS1081和nrdZ基因靶点，对170例结核性脑膜炎（确诊结核性脑膜炎（n = 146）、疑似结核性脑膜炎（n = 24））患者脑脊液样本（CSF）和100例非结核性脑膜炎对照进行Cf-DNA rt- pcr检测。还对Truenat MTB Plus试验和GeneXpert MTB/RIF Ultra进行了性能评估。结果：Cf-DNA M-rtPCR对TBM的总体诊断敏感性为78.24%，特异性为100%，阳性预测值为100%，阴性预测值为72.99%；对TBM的“明确”诊断敏感性为82.88%，特异性为100%，阳性预测值为100%，阴性预测值为80%。Cf-DNA M-rtPCR检测到Truenat MTB Plus试验和GeneXpert MTB/RIF Ultra未检测到的另外14例病例。在所使用的不同基因靶点中，仅IS1081检测到8例，仅IS6110检测到3例，仅nrdZ检测到1例。Truenat MTB Plus试验和GeneXpert MTB/RIF Ultra均报告6例对利福平耐药，并通过rpoB测序证实。结论：游离DNA检测结合m -rt - pcr可作为直接从脑脊液中诊断结核性脑膜炎的可靠工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Tuberculosis 医学-呼吸系统

CiteScore

4.60

自引率

3.10%

发文量

审稿时长

49 days

期刊介绍： Tuberculosis is a speciality journal focusing on basic experimental research on tuberculosis, notably on bacteriological, immunological and pathogenesis aspects of the disease. The journal publishes original research and reviews on the host response and immunology of tuberculosis and the molecular biology, genetics and physiology of the organism, however discourages submissions with a meta-analytical focus (for example, articles based on searches of published articles in public electronic databases, especially where there is lack of evidence of the personal involvement of authors in the generation of such material). We do not publish Clinical Case-Studies. Areas on which submissions are welcomed include: -Clinical TrialsDiagnostics- Antimicrobial resistance- Immunology- Leprosy- Microbiology, including microbial physiology- Molecular epidemiology- Non-tuberculous Mycobacteria- Pathogenesis- Pathology- Vaccine development. This Journal does not accept case-reports. The resurgence of interest in tuberculosis has accelerated the pace of relevant research and Tuberculosis has grown with it, as the only journal dedicated to experimental biomedical research in tuberculosis.