Publication: One-tube multiplex PCR method for rapid identification of Mycobacterium tuberculosis
3
Issued Date
2006-12-01
Resource Type
ISSN
01251562
Other identifier(s)
2-s2.0-33845749105
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Mahidol University
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SCOPUS
Bibliographic Citation
Southeast Asian Journal of Tropical Medicine and Public Health. Vol.37, No.3 (2006), 494-502
Suggested Citation
Angkana Chaiprasert, Therdsak Prammananan, Nipa Tingtoy, Preeyawis Na-Ubol, Somboon Srimuang, Kittipan Samerpitak, Watcharin Rangsipanuratn One-tube multiplex PCR method for rapid identification of Mycobacterium tuberculosis. Southeast Asian Journal of Tropical Medicine and Public Health. Vol.37, No.3 (2006), 494-502. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/23465
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Title
One-tube multiplex PCR method for rapid identification of Mycobacterium tuberculosis
Abstract
A rapid, inexpensive, simple, and accurate multiplex polymerase chain reaction (PCR) was developed in a single tube for identification of Mycobacterium tuberculosis. Assessment of sensitivity and specificity of simple PCR was performed with 116 strains of M. tuberculosis complex (MTC) and 144 strains of nontuberculous mycobacteria (NTM) compared with the biochemical method. Specific amplification of KS4, MTC-specific DNA fragment, was found in 98% (114/116) of MTC and not detected in 99% (143/144) of NTM. Amplification of the mtp40 gene revealed 95% sensitivity (100/105 strains of M. tuberculosis) and 77% specificity (not found in 119/155 mycobacterial strains). A multiplex PCR method based on the combination of KS4- and mrp40-derived primers was used for identification of M. tuberculosis. Crude DNA from slow growing mycobacteria with cream rough colonies that showed both 768-bp amplified product for KS4 and 396-bp for mtp40 was identified as M. tuberculosis whereas that from MTC gave only the 768-bp product.