Rapid identification of mycobacterium species by amplification of rpoB gene and reverse dot blot hybridization assay

Abstract

Abstract: Objective To establish a rapid,sensitive and specific method for identification of mycobacterium species through rpoB gene. Method Based on the sequence of rpoB gene,the standard strains of 24 Mycobacteria and 8 nonmycobacteria,37 clinical isolates of mycobacteria were detected by PCRreverse dot blot hybridization assay. Results 360 bp DNA fragments were amplified from all mycobacterial strains tested and were not found in all nonmycobacterial strains besides Hemolytic streptococcus and Corynebacterium pseudodiphtheriticum. 1 pg of M.tuberculosis DNA could be detected by PCR assay.Among 21 oligonucleotide probes,probe-M,fortuitum cross-hybridized with M.marinum,the other probes were specific.The Results of clinical isolates identified by this assay were in agreement with those obtained by conventional methods. Conclusion RpoB-based PCR-reverse dot blot hybridization assay might become a rapid,effective method for identification of mycobacterium species.

Key words: Mycobacterium, Polymerase Chain Reaction, Oligonucleotide Probes, Nucleic Acid Hybridization

Fan Bo,Wang Wei,Li Guoli,et al.. Rapid identification of mycobacterium species by amplification of rpoB gene and reverse dot blot hybridization assay[J]. Chinese Journal of Antituberculosis, 2006, 28(4): 212-215.

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