Application of nested-PCR technique for the diagnosis of tuberculosis

A test-system based on amplification of IS 986 fragment (nested-PCR) was developed for the detection of Mycobacterium tuberculosis and M. bovis in different biological samples. We constructed external primers and selected appropriate amplifications parameters (annealing temperatures for states I and II, the number of cycles for each amplification stage, components of the amplification mixture, and pretreatment conditions for different biological samples). The developed parameters make the detection of mycobacteria more efficient and less expensive compared to commercial Cobas Amplicor system.