Characterization of mycobacterial isolates from clinical samples by using PCR-RFLP assay targeting hsp65 gene region

Abstract

Mycobacterial have long been recognized as important human and animal pathogens. Mycobacterial diseases such as tuberculosis and leprosy continue to be an important public health problem all over the world. Not only M.tuberculosis and M. leprae are important one, but mycobacteria other than tuberculosis (MOTT) are also pathogens. The immunosuppressed individual infection by HIV infection have become the most significant risk factor for disseminated NTM diseases and of these 95% are due to Mycobacterial avium complex. Thus the rapid and specific diagnosis of tuberculosis is one of most pressing needs in effort to temporary treat and control the disease. Classical identification result for mycobacteria based on culture and biochemical test may take several weeks and test fail sometimes to produce precise identification. PCR- restriction analysis (PRA) is a PCR-RFLP based, very efficient and promising approach for species identification as it gives result within one day. Several targets for PRA analysis of mycobacteria’s are available which includes hsp 65kDa. The 65kDa proteins contains epitopes that are unique as well as epitopes that are common to various species of mycobacteria’s.PRA is based on amplification of 439bp fragment of hsp65 gene present in all mycobacteria offers an easy rapid an inexpensive procedure to identify several mycobacterial species in single experiment. Thus the current work emphasis on quick, advanced and robust methodology for diagnosis of tuberculosis and other mycobacterial diseases.