Journal of Biotechnology & Biomaterials
Open Access

Abstract

Tuberculosis remains a major public health problem worldwide with a global mortality of 1.5 million in 2014. Though early diagnosis is mandatory in control of TB especially for pulmonary TB as it is transmissible, it has remained enigmatic. A major hindrance to the diagnosis of EPTB is the atypical presentation, often simulating neoplasia and or inflammatory disorders. The conventional techniques used in TB diagnosis like AFB smear microscopy lack sensitivity and the gold standard, culture test takes time. The sensitivity and specificity were compared with AFB smear examination, Lowenstein-Jensen culture test and single step PCR. In order to find a sensitive and rapid technique nested multiplex PCR (nMPCR) targeting the IS6110 and MTP40 gene of Mycobacterium tuberculosis was evaluated for detection of M. tuberculosis DNA directly from clinical specimens of pulmonary and extra-pulmonary origin. A total of 200 clinical specimens from clinically suspected cases of extra-pulmonary tuberculosis and 20 control specimens of non-tuberculous aetiology were processed by smear, culture, single step and by nested multiplex PCR technique for detection of M. tuberculosis. The conventional culture was positive only in 150 (75%) of 200 specimens and 162 (81%) were single step PCR positive. The overall positivity of nested multiplex PCR was 100% (200/200). All the 20 control specimens were negative by nested multiplex PCR. Nested multiplex PCR increased the sensitivity of PCR and will be useful in diagnosing smear negative samples. Further, it can also be used to detect samples with M. tuberculosis strains lacking IS6110.

Biography

Email: pallavisinhamicro@gmail.com