ArticleJournal of clinical microbiology2010
Development and application of multiprobe real-time PCR method targeting the hsp65 gene for differentiation of Mycobacterium species from isolates and sputum specimens.
Article in Journal of clinical microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 42 citations in OpenAlex.
- Supporting Patients with Nontuberculous Mycobacterial Pulmonary Disease: Ensuring Best Practice in UK Healthcare Settings.Pharmacy (Basel, Switzerland) · 2024Review
- Molecular assessment of mycobacterial burden in the treatment of nontuberculous mycobacterial disease.ERJ open research · 2023Article
- Detection of non-tuberculosus mycobacteria (NTMs) in lung samples using 16S rRNA.Memorias do Instituto Oswaldo Cruz · 2022Article
- Identification of nontuberculous mycobacteria species by multiplex real-time PCR with high-resolution melting.Revista da Sociedade Brasileira de Medicina Tropical · 2020Article
- Development and evaluation of an in-house single step loop-mediated isothermal amplification (SS-LAMP) assay for the detection of Mycobacterium tuberculosis complex in sputum samples from Moroccan patients.BMC infectious diseases · 2016Article
- Culture-Independent Detection of Nontuberculous Mycobacteria in Clinical Respiratory Samples.Journal of clinical microbiology · 2016Article
- Separation of Mycobacterium abscessus into subspecies or genotype level by direct application of peptide nucleic acid multi-probe- real-time PCR method into sputa samples.BMC infectious diseases · 2015Article
- Evaluation of performance of the Real-Q NTM-ID kit for rapid identification of eight nontuberculous mycobacterial species.Journal of clinical microbiology · 2014Article
- Evaluation of hsp65 nested PCR-restriction analysis (PRA) for diagnosing tuberculosis in a high burden country.BioMed research international · 2013Article
- Evaluation of peptide nucleic acid probe-based real-time PCR for detection of Mycobacterium tuberculosis complex and nontuberculous mycobacteria in respiratory specimens.Annals of laboratory medicine · 2012Article
- Multiplex real-time PCR assay and melting curve analysis for identifying Mycobacterium tuberculosis complex and nontuberculous mycobacteria.Journal of clinical microbiology · 2012Article
- Discovery of a novel hsp65 genotype within Mycobacterium massiliense associated with the rough colony morphology.PloS one · 2012Article
- Comparison of a DNA Based PCR Approach with Conventional Methods for the Detection of Mycobacterium tuberculosis in Morocco.Mediterranean journal of hematology and infectious diseases · 2012Article
- First report of disseminated Mycobacterium skin infections in two liver transplant recipients and rapid diagnosis by hsp65 gene sequencing.Journal of clinical microbiology · 2011Article
- An appraisal of PCR-based technology in the detection of Mycobacterium tuberculosis.Molecular diagnosis & therapy · 2011Review
- Quantitative real-time PCR (q-PCR) for sputum smear diagnosis of pulmonary tuberculosis among people with HIV/AIDS.Revista do Instituto de Medicina Tropical de Sao PauloArticle
Corrections and comments
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Authors and funding
13 authors at 6 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We developed a multiprobe real-time PCR assay targeting hsp65 (HMPRT-PCR) to detect and identify mycobacterial isolates and isolates directly from sputum specimens. Primers and probes for HMPRT-PCR were designed on the basis of the hsp65 gene sequence, enabling the recognition of seven pathogenic mycobacteria, including Mycobacterium tuberculosis, M. avium, M. intracellulare, M. kansasii, M. abscessus, M. massiliense, and M. fortuitum. This technique was applied to 24 reference and 133 clinical isolates and differentiated between all strains with 100% sensitivity and specificity. Furthermore, this method was applied to sputum specimens from 117 consecutive smear-positive patients with smear results of from a trace to 3+. These results were then compared to those obtained using the rpoB PCR-restriction analysis method with samples from cultures of the same sputum specimens. The HMPRT-PCR method correctly identified the mycobacteria in 89 samples (76.0%, 89/117), and moreover, the sensitivity level was increased to 94.3% (50/53) for sputa with an acid-fast bacillus score equal to or greater than 2+. Our data suggest that this novel HMPRT-PCR method could be a promising approach for detecting pathogenic mycobacterial species from sputum samples and culture isolates routinely in a clinical setting.
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