Identification of Nontuberculous Mycobacterium Species by Polymerase Chain Reaction - Restriction Enzyme Analysis (PCR-REA) of rpoB gene in Clinical Isolates.

IF 1.6 Q4 INFECTIOUS DISEASES International Journal of Mycobacteriology Pub Date : 2024-07-01 Epub Date: 2024-09-14 DOI:10.4103/ijmy.ijmy_134_24
Raj Narayan Yadav, Yellanki Yashwanth Chowdary, Manpreet Bhalla, Ajoy Kumar Verma
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Abstract

Background: Nontuberculous mycobacteria (NTM) infections are an emerging global health concern with increasing incidence. Conventional identification methods for NTM species in clinical settings are prone to errors. This study evaluates a newer method, polymerase chain reaction-restriction enzyme analysis (PCR-REA) of the rpoB gene, for NTM species identification. The study identified NTM species in clinical samples using conventional biochemical techniques and compared the results with PCR-REA of the rpoB gene. This cross-sectional study was conducted at a tertiary health-care center in North India over 18 months, analyzing both pulmonary and extrapulmonary samples.

Methods: Two hundred and forty-seven NTM isolates were identified using phenotypic and biochemical methods. The same isolates were subjected to rpoB gene amplification by PCR followed by REA using Msp I and Hae III enzymes.

Results: Conventional methods identified 12 different NTM species (153 slow-growing and 94 rapid-growing), whereas PCR-REA identified 16 species (140 slow-growing, 107 rapid-growing). The Mycobacterium avium intracellulare complex was the most common species isolated. PCR-REA demonstrated higher resolution in species identification, particularly in differentiating within species complexes.

Conclusions: PCR-REA of the rpoB gene proves to be a simple, rapid, and more discriminative tool for NTM species identification compared to conventional methods. This technique could significantly improve the diagnosis and management of emerging NTM infections in clinical settings.

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通过聚合酶链式反应--rpoB 基因的限制酶分析(PCR-REA)鉴定临床分枝杆菌中的非结核分枝杆菌。
背景:非结核分枝杆菌(NTM)感染是一个新出现的全球健康问题,发病率不断上升。在临床环境中,NTM 物种的传统鉴定方法容易出错。本研究评估了一种用于鉴定 NTM 物种的较新方法,即 rpoB 基因的聚合酶链反应-限制酶分析(PCR-REA)。研究采用传统生化技术鉴定了临床样本中的 NTM 物种,并将结果与 rpoB 基因的 PCR-REA 进行了比较。这项横断面研究在印度北部的一家三级医疗保健中心进行,历时 18 个月,分析了肺部和肺外样本:方法:采用表型和生化方法鉴定了 247 株 NTM 分离物。这些分离物通过 PCR 进行 rpoB 基因扩增,然后使用 Msp I 和 Hae III 酶进行 REA:结果:传统方法鉴定出 12 种不同的非结核分枝杆菌(153 种生长缓慢,94 种生长迅速),而 PCR-REA 鉴定出 16 种(140 种生长缓慢,107 种生长迅速)。细胞内分枝杆菌复合体是最常见的分离菌种。PCR-REA 在物种鉴定方面表现出更高的分辨率,尤其是在物种复合体内部的区分方面:结论:与传统方法相比,rpoB 基因的 PCR-REA 被证明是一种简单、快速且更具鉴别力的非结核分枝杆菌物种鉴定工具。这项技术可大大改善临床环境中对新出现的 NTM 感染的诊断和管理。
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来源期刊
CiteScore
2.20
自引率
25.00%
发文量
62
审稿时长
7 weeks
期刊最新文献
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