Rapid Detection of Viable M. tuberculosis Directly from Sputum by Reverse Transcriptase PCR Targeting 85B Gene.
Loading...
Date
2011-06
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
This study was carried out to standardize reverse transcriptase PCR (RTPCR)
targeting 85B gene for the rapid detection of viable Mycobacterium
tuberculosis (M tuberculosis) from sputum specimens. 100 sputum
samples from clinically suspected tuberculosis patients were included in
the study. The sputum samples were collected in sterile diethyl
pyrocarbonate (DEPC) treated containers and transported in ice to the
laboratory within 2 hours to prevent degradation of RNA. The following
microbiological analysis was performed on the sputum specimens: Ziehl
Neelsen staining, Mycobacterial culture by BACTEC TB 460 reader and
RT-PCR targeting 85B gene. Out of the 100 sputum samples, 40 sputum
samples were Ziehl Neelsen stain positive, 58 sputum samples were
culture and RT-PCR positive and 42 were culture and RT-PCR were
negative. Among direct smear positive specimens, 3 specimens did not
grow in culture and was RT-PCR negative indicating the non-viability of
the acid-fast bacilli seen in the direct smear. The results of RT-PCR and
culture against control group and clinically diagnosed tuberculosis
patients were statistically significant by Chi square test (P<0.001).
RT-PCR targeting 85B gene of M tuberculosis standardized in our
laboratory for is a rapid, reliable and sensitive technique to detect the
viable M tuberculosis directly from sputum specimens.
Description
Keywords
M tuberculosis, Viable, RT-PCR
Citation
Ramasubban Gayathri, Kulandai Lily Therese, Sridhar R, Durai Kannan, Hajib N Madhavan, Meenakshi N. Rapid Detection of Viable M. tuberculosis Directly from Sputum by Reverse Transcriptase PCR Targeting 85B Gene. Journal of Communicable Diseases. 2011 Jun; 43(2): 89-96.