The polymerase chain reaction (PCR) is a rapid, reproducible and reliable method of detecting specific sequences of DNA. The sensitivity of PCR to various strains of mycobacterium depends on the degree to which the target sequence is conversed in the organism. The specificity depends on the extent to which the target sequence is unique to the organism. The objective of the study is to develop a rapid and specific laboratory test for detecting and identifying mycobacteria within clinical samples from nonhuman primates. In order to detect and differentiate the major pathogenic mycobacterial species, we used the sequence of published genup-specific primers to synthesize probes. This was done in order to amplify the dnaJ genes from a broad spectrum of mycobacterial species. We used dot blot hybridization analysis with species-specific oligonucleotide probes for the specific nonhuman primate mycobacterium, allowing a rapid identifiction of these species follow polymerase chain reaction of the dnaJ gene. Presently, results indicate that the polymerase chain reaction with genus-specific oligonucleotide probes is useful for diagnosis of nonhuman primate mycobacterial infection.