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Research article
PCR-reverse blot hybridization assay in respiratory specimens for rapid detection and differentiation of mycobacteria in HIV-negative population
He-ping Xiao
Tongji University Affiliated Shanghai Pulmonary Hospital
Corresponding Author
ORCiD: https://orcid.org/0000-0002-9186-6613
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Objective: Rapid identification of pathogenic Mycobacterium species is critical for a successful treatment. However, traditional method is time-consuming and cannot discriminate isolated NTM to species level. In the retrospective study, we evaluated the clinical applicability of PCR-reverse blot hybridization assay (PCR-REBA) 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 and 210 patients with non-mycobacterium pulmonary disease used as controls were analyzed. Sputum samples or BALF were obtained for MGIT 960 and PCR-REBA. To confirm mycobacterium species inconsistently identified by the two different assays, high resolution melting (HRM) analysis was performed.
Results: A total of 334 sputum and 362 BALF specimens from 696 patients with mycobacterium pulmonary disease (292 MTB and 404 NTM) were eventually analyzed. In total, 292 MTBC and 436 NTM isolates (co-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%), M. abscessus (n=106, 24.3%), M. kansasii (n=46, 10.6%), M. avium (n=36, 8.3%). Twenty-two cases had mixed infection with both M. intracellulare and M. abscessus and ten cases had mixed infection with both M. avium and M. abscessus. A high level of agreement (n=696; 94.5%) was found between MGIT 960 and PCR-REBA (k = 0.845, P = 0.000). PCR-REBA had the higher AUC than MGIT 960 for both MTBC and NTM.
Conclusion: PCR-REBA is helpful for rapid mycobacterial species identification with low cost and simplicity and therefore recommending its routine use.
Keywords
Mycobacterium tuberculosis (MTB), nontuberculous mycobacteria (NTM), identification, PCR-reverse blot hybridization assay (PCR-REBA), molecular diagnosis
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This preprint is under consideration at BMC Infectious Diseases. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
PCR-reverse blot hybridization assay in respiratory specimens for rapid detection and differentiation of mycobacteria in HIV-negative population
He-ping Xiao
Tongji University Affiliated Shanghai Pulmonary Hospital
Corresponding Author
ORCiD: https://orcid.org/0000-0002-9186-6613
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 28 May, 2020
No community comments so far
Editor assigned
On 20 May, 2020
Submission checks complete
On 19 May, 2020
Editor invited
On 19 May, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Infectious Diseases
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Objective: Rapid identification of pathogenic Mycobacterium species is critical for a successful treatment. However, traditional method is time-consuming and cannot discriminate isolated NTM to species level. In the retrospective study, we evaluated the clinical applicability of PCR-reverse blot hybridization assay (PCR-REBA) 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 and 210 patients with non-mycobacterium pulmonary disease used as controls were analyzed. Sputum samples or BALF were obtained for MGIT 960 and PCR-REBA. To confirm mycobacterium species inconsistently identified by the two different assays, high resolution melting (HRM) analysis was performed.
Results: A total of 334 sputum and 362 BALF specimens from 696 patients with mycobacterium pulmonary disease (292 MTB and 404 NTM) were eventually analyzed. In total, 292 MTBC and 436 NTM isolates (co-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%), M. abscessus (n=106, 24.3%), M. kansasii (n=46, 10.6%), M. avium (n=36, 8.3%). Twenty-two cases had mixed infection with both M. intracellulare and M. abscessus and ten cases had mixed infection with both M. avium and M. abscessus. A high level of agreement (n=696; 94.5%) was found between MGIT 960 and PCR-REBA (k = 0.845, P = 0.000). PCR-REBA had the higher AUC than MGIT 960 for both MTBC and NTM.
Conclusion: PCR-REBA is helpful for rapid mycobacterial species identification with low cost and simplicity and therefore recommending its routine use.
Figure 1
Figure 2
Figure 3