Rapid diagnosis of oral tuberculosis by amplification of Mycobacterium DNA from paraffin embedded specimens.

Abstract

The polymerase chain reaction (PCR) assay is a powerful tool for quick diagnosis of various infectious diseases. We applied this technique as well as conventional histopathological examination to diagnose oral tuberculosis. Ziehl-Neelsen staining of oral mucosal specimens often fails to detect Mycobacterium (M.) tuberculosis due to the low number of bacteria in the tissue. Specific primers and probes were synthesized based upon the nucleotide sequence of the 65 kDa membrane protein of M. tuberculosis. DNA extracted from the paraffin-embedded tissue was amplified using taq polymerase. PCR assay detected M. tuberculosis in 5 of 6 samples. Although the gene segments from these species were quite similar, the gamma 32P labeled noligonucleotide probes distinguished between M. tuberculosis and M. fotuitum by southern blot hybridization. In all specimens that were Ziehl-Neelsen negative, M. tuberculosis DNA was detected by PCR. These results suggest that PCR is a useful means of diagnosing mycobacterium infection.