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Polymerase Chain Reaction using the MPB64 fragment for detection of mycobacterium tuberculosis complex DNA in suspected TB cases

Ritambhara Aryal, Anil K. Sah, Deepak S. Paudel, Bishnu Joshi, Sunil P. Lekhak, Reema Rajbhandari, Bal Hari Poudel, Bijaya Pant, Ganesh Prasad Neupane, Basant Pant.




Abstract

Background: Various primers have been used for detection of Mycobacterium tuberculosis complex (MTBC) DNA using Polymerase chain reaction (PCR). The MPB64 gene has been demonstrated to be a highly specific target for MTBC detection.

Aims & Objectives: To detect MTBC DNA using MPB64 primers in clinical specimens of Nepalese patients suspected to have tuberculosis (TB).

Materials and Methods: DNA was extracted from 44 clinical specimens of patients suspected of having TB and MPB64 targeted PCR was performed.

Results: Bands comprising 240 base pair (bp) of MPB64 sequence were present in 15 of the 44 clinical samples, of which 4 were pulmonary and the remainders were extra-pulmonary samples. The overall positivity of MPB64 targeted PCR was 34.1%.

Conclusion: PCR targeting the MPB64 fragment has a potential of detecting the MTBC DNA and has potentially valuable clinical applications in early detection of TB in Nepal.

Key words: MPB64; Tuberculosis; Extra- Pulmonary Tuberculosis; Polymerase Chain Reaction






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