[Rapid identification of bacteria by PCR and hybridization].
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.
Ishiko
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Mitsubishi-Yuka Bio-Clinical Laboratories Inc., Division of Molecular Diagnosis of Infectious Disease.
D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis*; D016480:Helicobacter pylori / Q000235:genetics / Q000302:isolation & purification*; D016106:Methicillin Resistance; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification*
D004269:DNA, Bacterial