Journal Article

Comparison of molecular testing methods for diagnosing non-tuberculous mycobacterial infections

Pan, Jue, Fu, Aisi, Miao, Qing, Yao, Yumeng, Li, Na, Yin, Xiaoyu, Pang, Bin, Qin, Le, Chen, Yu, Luan, Sichun, Jin, Wenting, Bao, Rong, Wang, Qingqing, Zhou, Chunmei, Hu, Bijie, Ji, Yuan, Fang, Tingting, Wang, Leilei, Zhu, Pengyan

2025

Overview

Purposes Rapid and accurate identification of non-tuberculous mycobacteria (NTM) is crucial yet challenging, promoting the development of novel molecular techniques such as amplification-based targeted high-throughput sequencing and metagenomic unbiased high-throughput sequencing. We aimed to evaluate the diagnostic value of these molecular techniques for NTM infection. Methods A total of 115 clinical specimens from patients with confirmed NTM infection were subjected to multiplex polymerase chain reaction detection techniques (multi-PCR), metagenomic Next-Generation Sequencing (mNGS), targeted Next-Generation Sequencing (tNGS), and targeted Nanopore sequencing (tNanopore). Positivity rates and species identification were compared among these techniques. Results The sensitivity of mNGS, tNGS, and multi-PCR in NTM-infection diagnosis was 44.3%, 42.6%, and 36.5%, respectively, while the sensitivity of the three methods in combination increased to 54.8%. The pathogen identification results of mNGS, tNGS and multi-PCR were matched in 80.6% (25/31) samples at the species level, among which 14 samples (45.2%) was completely matched at the subspecies level. The results of tNanopore, tNGS and mNGS at the species level were completely matched in 73.3% (22/30) samples. Conclusions These molecular assays demonstrated comparable performance in precisely identifying NTM species in clinical specimens, showing their promising potential as efficient and alternative tools for the rapid diagnosis of NTM disease.

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V
• Subject: Adult / Aged / Aged, 80 and over / Biomedical and Life Sciences / Biomedicine / Diagnosis / Female / High-Throughput Nucleotide Sequencing - methods / Humans / Infections / Internal Medicine / Male / Medical Microbiology / Metagenomics / Middle Aged / Molecular chains / Molecular Diagnostic Techniques - methods / Multiplex Polymerase Chain Reaction - methods / Mycobacterium Infections, Nontuberculous - diagnosis / Mycobacterium Infections, Nontuberculous - microbiology / Nanopore Sequencing / Next-generation sequencing / Nontuberculous Mycobacteria - classification / Nontuberculous Mycobacteria - genetics / Nontuberculous Mycobacteria - isolation & purification / Polymerase chain reaction / Samples / Sensitivity / Sensitivity and Specificity / Tuberculosis