1887

Abstract

For the rapid identification of , multiplex PCR, using 16S rDNA as target gene, was used and its value was evaluated. Forward primers specific for and , among the five validly described species, were designed from the alignment of 16S rDNA sequences. has been shown to be a member of the same species as by recent molecular studies and preliminary DNA-DNA relatedness tests. Therefore, and were considered as members of the same species in this study. Each primer was found to be species-specific by specificity testing. NSP01, NSP19, NSP21and NSP22could be clearly differentiated by PCR products characteristic for each species in the multiplex PCR assay. gave an identical result to NSP01. The additional 17 strains of and the additional four strains of gave PCR products identical to those of NSP01and NSP22, respectively. Multiplex PCR was found to be rapid, species-specific and reproducible. The technique evaluated in this study proved to be effective for rapidly identifying strains to species level.

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1998-07-01
2022-05-28
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References

  1. Bourque S. N., Valero J. R., Mercier J., Lavoie M. C., Levesque R. C. 1993; Multiplex polymerase chain reaction for detection and differentiation of the microbial insecticide Bacillus thurin-giensis. Appl Environ Microbiol 59:523–527
    [Google Scholar]
  2. Collins M. D., Stackebrandt E. 1989; Molecular taxonomic studies on some LL-diaminopimelic acid-containing coryne- forms from herbage: description of Nocardioides fastidiosa sp. nov. FEMS Microbiol Lett 57:289–294
    [Google Scholar]
  3. Collins M. D., Dorsch M., Stackebrandt E. 1989; Transfer of Pimelobacter tumescens to Terrabacter gen. nov. as Terrabacter tumescens comb. nov. and of Pimelobacter jensenii to Nocardioides as Nocardioides jensenii comb. nov. IntJSyst Bacteriol 39:1–6
    [Google Scholar]
  4. Collins M. D., Cockcroft S., Wallbanks S. 1994; Phylogenetic analysis of a new LL-diaminopimelic acid-containing coryne- form bacterium from herbage, Nocardioidesplantarum sp. nov. Int J Syst Bacteriol 44:523–526
    [Google Scholar]
  5. Frothingham R., Allen R. L., Wilson K. H. 1991; Rapid 16S ribosomal DNA sequencing from a single colony without DNA extraction or purification. Biotechniques 11:42–44
    [Google Scholar]
  6. Geha D. J., Uhl J. R., Gustaferro C. A., Persing D. H. 1994; Multiplex PCR for identification of methicillin-resistant staphylococci in the clinical laboratory. J Clin Microbiol 32:1768–1772
    [Google Scholar]
  7. Hiraishi A., Shin Y. K., Ueda Y., Sugiyama J. 1994; Automated sequencing of PCR-amplified 16S rDNA on ‘ Hydrolink ’ gels. J Microbiol Methods 19:145–154
    [Google Scholar]
  8. Laguerre G., Allard M.-R., Revoy F., Amarger N. 1994; Rapid identification of rhizobia by restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA genes. Appl Environ Microbiol 60:56–63
    [Google Scholar]
  9. Miller E. S., Woese C. R., Brenner S. 1991; Description of the erythromycin-producing bacterium Arthrobacter sp. strain NRRL B-3381 as Aeromicrobium erythreum gen. nov., sp. nov. Int J Syst Bacteriol 41:363–368
    [Google Scholar]
  10. O'Donnell A. G., Goodfellow M., Minnikin D. E. 1982; Lipids in the classification of Nocardioides'. reclassification of Arthrobacter simplex (Jensen) Lochhead in the genus Nocardioides (Prauser) emend. O’Donnell et al. as Nocardioides simplex comb. nov. Arch Microbiol 133:323–329
    [Google Scholar]
  11. Persing D. H., Smith T. F., Tenover F. C., White T. J. 1993 Diagnostic Molecular Microbiology: Principles and Applications Washington, DC: American Society for Microbiology;
    [Google Scholar]
  12. Prauser H. 1976; Nocardioides, a new genus of the order Actinomycetales. Int J Syst Bacteriol 26:58–65
    [Google Scholar]
  13. Prauser H. 1984; Nocardioides luteus sp. nov. Z Allg Microbiol 24:647–648
    [Google Scholar]
  14. Stackebrandt E., Rainey F. A., Ward-Rainey N. L. 1997; Proposal for a new hierarchic classification system, Actino- bacteria classis nov. Int J Syst Bacteriol 47:479–491
    [Google Scholar]
  15. Suzuki K.-I., Komagata K. 1983; Pimelobacter gen. nov., a new genus of coryneform bacteria with LL-diaminopimelic acid in the cell wall. J Gen Appl Microbiol 29:59–71
    [Google Scholar]
  16. Tamura T., Yokota A. 1994; Transfer of Nocardioides fastidiosa Collins and Stackebrandt 1989 to the genus Aeromicrobium as Aeromicrobium fastidiosum comb. nov. Int J Syst Bacteriol 44:608–611
    [Google Scholar]
  17. Vandamme P., Pot B., Gillis M., De Vos P., Kersters K., Swings J. 1996; Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiol Rev 60:407–438
    [Google Scholar]
  18. Wayne L. G., Brenner D. J., Colwell R. R., Grimont P. A. D., Kandler O., Krichevsky M. I., Moore L. H., Moore W. E. C., Murray R. G. E., Stackebrandt E., Starr M. P., TrUper H. G. 1987; Report of the ad hoc committee on reconciliation of approaches to bacterial systematics. Int J Syst Bacteriol 37:463–464
    [Google Scholar]
  19. Welsh J., McClelland M. 1990; Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18:7213–7218
    [Google Scholar]
  20. Williams J. G. K., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. 1990; DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535
    [Google Scholar]
  21. Woese C. R. 1987; Bacterial evolution. Microbiol Rev 51:221–271
    [Google Scholar]
  22. Yoon J.-H., Kim H., Kim S.-B., Kim H.-J., Kim W. Y., Lee S. T., Goodfellow M., Park Y.-H. 1996a; Identification of Saccharo- monospora strains by the use of genomic DNA fragments and rRNA gene probes. Int J Syst Bacteriol 46:502–505
    [Google Scholar]
  23. Yoon J.-H., Lee S. T., Shin Y. K., Kim S.-B., Kim H.-J., Goodfellow M., Park Y.-H. 1996b; Rapid identification of Saccharomonospora strains by multiplex PCR using species- specific primers within the 16S rRNA gene. J Microbiol Methods 27:89–95
    [Google Scholar]
  24. Yoon J.-H., Lee S. T., Kim S.-B., Kim W. Y., Goodfellow M., Park Y.-H. 1997a; Restriction fragment length polymorphism analysis of PCR-amplified 16S ribosomal DNA for rapid identification of Saccharomonospora strains. IntJSyst Bacteriol 47:111–114
    [Google Scholar]
  25. Yoon J.-H., Lee J.-S., Shin Y. K., Park Y.-H., Lee S. T. 1997b; Reclassification of Nocardioides simplex ATCC 13260, ATCC 19565 and ATCC 19566 as Rhodococcus erythropolis. Int J Syst Bacteriol 47:904–907
    [Google Scholar]
  26. Yoon J.-H., Lee S. T., Park Y.-H. 1998a; Inter-and intraspecific phylogenetic analysis of the genus Nocardioides and related taxa based on 16S rDNA sequences. Int J Syst Bacteriol 48:187–194
    [Google Scholar]
  27. Yoon J.-H., Lee S. T., Park Y.-H. 1998b; Genetic analyses of the genus Nocardioides and related taxa based on 16S-23S rDNA internally transcribed spacer sequences. Int J Syst Bacteriol 48:641–650
    [Google Scholar]
  28. Zabeau M., Vos P. 1993 Selective restriction fragment amplification: a general method for DNA fingerprinting European Patent Office, patent no. 0534858 Al
    [Google Scholar]
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