Publication: Multi-probe real-time PCR identification of common mycobacterium species in blood culture broth
Issued Date
2009-01-01
Resource Type
ISSN
15251578
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2-s2.0-58849118489
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Molecular Diagnostics. Vol.11, No.1 (2009), 42-48
Suggested Citation
Suporn Foongladda, Suporn Pholwat, Boonchuay Eampokalap, Pattarachai Kiratisin, Ruengpung Sutthent Multi-probe real-time PCR identification of common mycobacterium species in blood culture broth. Journal of Molecular Diagnostics. Vol.11, No.1 (2009), 42-48. doi:10.2353/jmoldx.2009.080081 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/27332
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Title
Multi-probe real-time PCR identification of common mycobacterium species in blood culture broth
Abstract
Mycobacterium tuberculosis complex, M. avium, and M. intraceliulare are the most common causes of systemic bacterial infection in AIDS patients. To identify these mycobacterial isolates in primary blood culture broths, we developed a multiple hybridization probe-based real-time PCR assay using the LightCycler system. The primers were designed to amplify a 320-bp fragment of Mycobacterium 16S rRNA genes. Reaction specificity was evaluated using PCR amplification curves along with specific melting temperatures of probes on DNA extracted from 13 Mycobacterium species. In this study, results showed 100% accuracy for the selected bacterial panel. Detection limits were 350, 600, and 650 colony-forming unit (CFU)/ml blood culture broths for M. tuberculosis complex, M. avium, and M. intracellular, respectively (1 to 2 CFU/reaction). To evaluate clinical applicability, 341 acid-fast bacilli in blood culture broths were analyzed. In total, 327 (96%) were positively identified: 54.5% M. tuberculosis complex, 37.5% M. avium, and 3.8% M. intracellulare. Results can be available within 3 hours of receiving a broth sample, which makes this rapid and simple assay an attractive diagnostic tool for clinical use. Copyright © American Society for Investigative Pathology and the Association for Molecular Pathology.