[Rapid detection and identification of mycobacterial DNA by PCR].
We have developed a highly sensitive and rapid PCR assay for detection and identification of mycobacterial species. Two sets of PCR primers were prepared; one was for the 19 kDa antigen gene and the other for the dna J gene. The PCR for 19 kDa antigen gene was found to be specific to the M. tuberculosis complex; M. tuberculosis, M. bovis and M. africanum. On the other hand, the PCR for the dna J gene was able to detect a broad spectrum of mycobacterial species including M. tuberculosis, M. avium, M. intracellulare and M. kansasii. The sensitivity of PCR was similar to that of the culture method for precultured M. tuberculosis whereas PCR was much more sensitive than the culture for clinical samples. The procedure of decontamination by sodium hydroxide for clinical samples could reduce the viability of M. tuberculosis while it did not affect mycobacterial DNA. The nucleotide sequence homologies among major mycobacteria were also determined following PCR for the dna J gene. This allowed us to make restriction maps for each mycobacterium. We have demonstrated that the PCR-RFLP for the dna J gene will be a useful laboratory test for rapid identification of tuberculous and nontuberculous mycobacterial species.
Nagai
R
R
Department of Laboratory Medicine, University of Tokyo.
Takewaki
S
S
Wada
A
A
Okuzumi
K
K
Tobita
A
A
Ohkubo
A
A
D001483:Base Sequence; D004269:DNA, Bacterial / Q000302:isolation & purification*; D005784:Gene Amplification; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
D004269:DNA, Bacterial