Species Specific DNA Profiling Mycobacterial Genomes Using Polymerase Chain Reaction with Single Universal Primer (UP-PCR)

Three tuberculous, twenty-one non-tuberculous mycobacterial (NTM) reference strains and seventy two isolates classified by biochemical tests were shown to produce specific sets of DNA fragments in a polymerase chain reaction with single universal primer (UP-PCR). A rather wide limit of tolerance for variations in procedure of PCR mixture preparation and thermocycling parameters was found. There was good correlation between UP-PCR pattern obtained in an agarose gel for each strain and the absolute majority of isolates tested. The PCR products of M. tuberculosis and M. bovis were shown to be very similar and well distinctive from that of all NTM strains tested. When image analysis system was applied to profile the UP-PCR patterns, all M. tuberculosis complex isolates were well differentiated from NTM species according to their specific profiles. Thus, UP-PCR profiling could give an efficient means for discriminating mycobacteria that utilizes the least time and applicable to controversial cases of suspected tuberculosis.