Rapid identification of bacteria by PCR and hybridization
Ishiko, H.
Nihon Rinsho. Japanese Journal of Clinical Medicine 52(2): 344-349
1994
ISSN/ISBN: 0047-1852 PMID: 8126883 Document Number: 425979
The amplification of bacteria DNA by PCR followed by rapid identification with hybridization are described. In the case of Mycobacteria, a 206 bases in dnaJ gene was amplified by nested PCR with conserved primers. The amplified DNAs were then hybridized with species-specific oligoprobes. Theses oligoprobes are capable of identifying the amplified DNA as M. tuberculosis, M. avium, M. intracellulare, M. kansasii or others. In the case of MRSA, clinical samples were examined to amplify mecA, femA and tsst-1 genes in the same tube, using mixed primers and the resultant 680 bp, 307 bp and 121 bp were identified by hybridization. For Helicobacter pylori, a 203 base were amplified and identified with [32p]-labeled oligoprobe as described (Valentine, J.L. et al., J. Clin. Microbiol., 29:689-695, 1991.). The sensitivity was 10(2) CFU/tube on agarose gel and 10 CFU/tube by hybridization.