Nucleic acid sequence-based amplification (NASBA) for the identification of mycobacteria

J Gen Microbiol. 1993 Oct;139(10):2423-9. doi: 10.1099/00221287-139-10-2423.


Nucleic acid sequence-based amplification (NASBA), an isothermal amplification technique for nucleic acids (NA), was investigated for the species-specific identification of mycobacteria. A set of primers was selected from a highly conserved region of the 16S rRNA sequence of mycobacteria sandwiching a variable sequence to perform amplification of mycobacterial RNA. Species-specific probes for the M. tuberculosis complex, M. avium-paratuberculosis, M. intracellulare and M. leprae were hybridized in-solution with the amplified nucleic acids of 10 pathogenic mycobacteria and 11 closely related bacteria, as well as with human-derived NA in an enzyme-linked gel assay (ELGA). Each probe was shown to hybridize specifically to the amplified single-stranded RNA of the corresponding species. Thirty-two clinical isolates of M. tuberculosis strains from different parts of the world were correctly identified by NASBA using the M. tuberculosis-complex-specific probe. In combination with the ELGA, NASBA could identify mycobacteria rapidly, i.e. in less than 6 h. The relative simplicity and rapidity of this technique makes it an attractive tool for species-specific identification of mycobacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Ribosomal / genetics
  • Molecular Sequence Data
  • Mycobacterium / genetics*
  • Mycobacterium / isolation & purification*
  • Mycobacterium avium / genetics
  • Mycobacterium avium / isolation & purification
  • Mycobacterium leprae / genetics
  • Mycobacterium leprae / isolation & purification
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / isolation & purification
  • Nucleic Acid Amplification Techniques*
  • RNA, Ribosomal, 16S / genetics
  • Sensitivity and Specificity
  • Species Specificity


  • DNA, Ribosomal
  • RNA, Ribosomal, 16S