聚合酶链反应-反向斑点杂交法在呼吸道标本中用于快速检测和区分 HIV 阴性人群中的分枝杆菌。
PCR-reverse blot hybridization assay in respiratory specimens for rapid detection and differentiation of mycobacteria in HIV-negative population.
机构信息
Clinic and Research Center of Tuberculosis, Department of Tuberculosis, Shanghai, Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China.
Shanghai Clinic and Research Center of Tuberculosis, Department of Tuberculosis, Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Tongji University School of Medicine, No. 507 Zhengmin Road, Shanghai, 200433, China.
出版信息
BMC Infect Dis. 2021 Mar 16;21(1):264. doi: 10.1186/s12879-021-05934-x.
BACKGROUND
Rapid identification of pathogenic Mycobacterium species is critical for a successful treatment. However, traditional method is time-consuming and cannot discriminate isolated non-tuberculosis mycobacteria (NTM) at species level. In the retrospective study, we evaluated the clinical applicability of PCR-reverse blot hybridization assay (PCR-REBA Myco-ID) with clinical specimens for rapid detection and differentiation of mycobacterial species.
METHODS
A total of 334 sputum and 362 bronchial alveolar lavage fluids (BALF) from 696 patients with mycobacterium pulmonary disease (MPD) and 210 patients with non-mycobacterium pulmonary disease used as controls were analyzed. Sputum or BALF were obtained for MGIT 960-TBc ID test and PCR-REBA Myco-ID assay. High resolution melt analysis (HRM) was used to resolve inconsistent results of MGIT 960-TBc ID test and PCR-REBA Myco-ID assay.
RESULTS
A total of 334 sputum and 362 BALF specimens from 696 MPD patients (292 MTB and 404 NTM) were eventually analyzed. In total, 292 MTBC and 436 NTM isolates (mixed infection of two species in 32 specimens) across 10 Mycobacterium species were identified. The most frequently isolated NTM species were M. intracellulare (n = 236, 54.1%), followed by M. abscessus (n = 106, 24.3%), M. kansasii (n = 46, 10.6%), M. avium (n = 36, 8.3%). Twenty-two cases had M. intracellulare and M. abscessus mixed infection and ten cases had M. avium and M. abscessus mixed infection. A high level of agreement (n = 696; 94.5%) was found between MGIT 960-TBc ID and PCR-REBA Myco-ID (k = 0.845, P = 0.000). PCR-REBA Myco-ID assay had higher AUC for both MTBC and NTM than MGIT 960-TBc ID test.
CONCLUSION
PCR-REBA Myco-ID has the advantages of rapid, comparatively easy to perform, relatively low cost and superior accuracy in mycobacterial species identification compared with MGIT 960-TBc ID. We recommend it into workflow of mycobacterial laboratories especially in source-limited countries.
背景
快速鉴定致病性分枝杆菌物种对于成功治疗至关重要。然而，传统方法耗时且无法在物种水平上区分分离的非结核分枝杆菌（NTM）。在回顾性研究中，我们评估了 PCR-反向斑点杂交检测（PCR-REBA Myco-ID）与临床标本联合用于快速检测和区分分枝杆菌物种的临床适用性。
方法
分析了 696 例分枝杆菌肺病（MPD）患者和 210 例非分枝杆菌肺病患者的 334 份痰和 362 份支气管肺泡灌洗液（BALF）。使用 MGIT 960-TBc ID 检测和 PCR-REBA Myco-ID 检测分析痰或 BALF。高分辨率熔解分析（HRM）用于解决 MGIT 960-TBc ID 检测和 PCR-REBA Myco-ID 检测结果不一致的问题。
结果
共分析了 696 例 MPD 患者（292 例 MTB 和 404 例 NTM）的 334 份痰和 362 份 BALF 标本。共鉴定了 10 种分枝杆菌种的 292 株 MTBC 和 436 株 NTM 分离株（32 份标本中有两种混合感染）。最常分离的 NTM 种是 M. intracellulare（n=236，54.1%），其次是 M. abscessus（n=106，24.3%）、M. kansasii（n=46，10.6%）、M. avium（n=36，8.3%）。22 例有 M. intracellulare 和 M. abscessus 混合感染，10 例有 M. avium 和 M. abscessus 混合感染。MGIT 960-TBc ID 和 PCR-REBA Myco-ID 之间有很高的一致性（n=696；94.5%）（k=0.845，P=0.000）。与 MGIT 960-TBc ID 检测相比，PCR-REBA Myco-ID 检测对 MTBC 和 NTM 的 AUC 值更高。
结论
与 MGIT 960-TBc ID 相比，PCR-REBA Myco-ID 具有快速、相对容易操作、成本相对较低、分枝杆菌物种鉴定准确性高的优点。我们建议将其纳入分枝杆菌实验室的工作流程，特别是在资源有限的国家。
Zotero 插件