Simple and rational approach to the identification of Mycobacterium tuberculosis, Mycobacterium avium complex species, and other commonly isolated mycobacteria.

A novel PCR-restriction fragment length polymorphism analysis of the hsp65 gene was developed. The restriction patterns for Mycobacterium tuberculosis and Mycobacterium avium complex (MAC) species were designed to be highly distinct, and the overall number of restriction patterns was designed to be limited. Four hundred specimens (17 reference strains and 383 clinical isolates) were tested, of which 98 were M. tuberculosis and 132 were MAC species. The assay was virtually 100% sensitive and specific for M. tuberculosis and MAC species. Moreover, it gave highly concordant results for other mycobacterial species other than M. terrae complex species. This assay can be completed in one day and is user-friendly and robust. Therefore, it is highly suitable for large-scale use in a clinical laboratory.