Abstract:
The present study was designed to demonstrate the molecular diagnosis usefulness for direct detection of M. tuberculosis from clinical samples. A method of Duplex Polymerase Chain Reaction (DPCR) and Real-time PCR (RTPCR) with primer specific for detection of the 156110 insertion element of M. tuberculosis complex and the p53gene of Human Beta Globin (Hbb) were developed. Fifty-four samples consisted of 25 from pulmonary and 29 from non-pulmonary specimens of patients suspected with M. tuberculosis infection. In a comparison experiment, 37 and 34 samples were detected as positive by ordinary and duplex PCR (DPCR), respectively, wherein data variation was detected in samples of pleural and other body fluids. For pulmonary specimens, 12/25 (46%) and 5/25 (19%) were positive as detected by DPCR and RTPCR, respectively. On the other hand, the number of positive results, 22/29 (71 %) and 8/29 (21 %) was higher among non-pulmonary specimens as detected by DPCR and RTPCR, respectively. Of these number, DPCR and RTPCR identified 15/34 (44%) and 8/34 (24%) of specimens, respectively in which Acid Fast Bacilli (AFB) and culture were negative. In addition, 18/19 (95%) and 4/19 (21 %) of the AFB positive specimens can be detected by DPCR and RTPCR, respectively. Nevertheless, both PCR amplification methods were able to amplify the 156110 genes for culture positive specimen. Therefore, duplex PCR method can be a useful additional technique for the diagnosis of M. tuberculosis suspected infection. However, RTPCR assay still need to be established.
Page(s):
100-105
DOI:
DOI not available
Published:
Journal: Biotechnology, Volume: 7, Issue: 1, Year: 2008