Publication: Detection of Mycobacterium tuberculosis from sputum specimens using one-tube nested PCR
Issued Date
2001-03-01
Resource Type
ISSN
01251562
Other identifier(s)
2-s2.0-0035288804
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Mahidol University
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SCOPUS
Bibliographic Citation
Southeast Asian Journal of Tropical Medicine and Public Health. Vol.32, No.1 (2001), 114-125
Suggested Citation
Nipa Gengvinij, Sa Nga Pattanakitsakul, Nitipatana Chierakul, Angkana Chaiprasert Detection of Mycobacterium tuberculosis from sputum specimens using one-tube nested PCR. Southeast Asian Journal of Tropical Medicine and Public Health. Vol.32, No.1 (2001), 114-125. Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/26838
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Title
Detection of Mycobacterium tuberculosis from sputum specimens using one-tube nested PCR
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Abstract
One-tube nested PCR was developed for diagnosis of pulmonary tuberculosis using sequences based on the 16SrRNA gene. The usage of primers 16SOL, 16SOR, 16SIL and 16SIR with optimized conditions could detect 555 bp DNA band from 21 species, 41 strains of mycobacteria and one isolate of Nocardia asteroides. It also revealed a specific 306 bp DNA band from 59 strains of M. tuberculosis complex. Cross amplification was observed in M. marinum, M. ulcerans and a few isolates of M. fortuitum complex. The developed method could detect as little as 100 fg of M. tuberculosis DNA. The PCR mixtures could be stored at 0°C for 2 months or at -20°C for at least 20 months without decrease in sensitivity. Using one-tube nested PCR for detection of M. tuberculosis compared with acid fast staining and culture results from 153 sputum specimens revealed 88.6% sensitivity and 89.2% specificity in smear positive specimens and 93.2% sensitivity and 85.0% specificity in culture positive specimens.