Abstract
BackgroundRapid 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.MethodsA 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.ResultsA 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.ConclusionPCR-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.
Highlights
Rapid identification of pathogenic Mycobacterium species is critical for a successful treatment
The genus Mycobacterium contains a number of acidfast bacilli (AFB), including Mycobacterium tuberculosis complex (MTBC), Mycobacterium leprae, and nontuberculosis mycobacteria (NTM) [1]
Demographic and clinical characteristics of the participants In all, 1378 patients (698 sputum and 680 bronchial alveolar lavage fluids (BALF)) with clinically suspected non-tuberculosis mycobacteria (NTM)-PD were enrolled into this study
Summary
Rapid identification of pathogenic Mycobacterium species is critical for a successful treatment. Traditional method is time-consuming and cannot discriminate isolated non-tuberculosis mycobacteria (NTM) at species level. The traditional methods used in mycobacterial laboratories require cultivated isolates and the results are only obtained after weeks to months of incubation. These culture-based methods are timeconsuming and labor-intensive [16]. The traditional methods could not discriminate NTM to the species level, or detect mixed infections in which two or more mycobacterial species were simultaneously detected in the same specimen [17]. IGRAs are based on M. tuberculosis specific antigens These antigens are not present in NTM strains other than M. marinum, M. kansasii, M. gordonae and M. szulgai [18]. IGRAs could not make a reliably distinction between NTM and MTBC
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