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. 1994 Mar;32(3):740–745. doi: 10.1128/jcm.32.3.740-745.1994

Rapid identification of mycolic acid patterns of mycobacteria by high-performance liquid chromatography using pattern recognition software and a Mycobacterium library.

S E Glickman 1, J O Kilburn 1, W R Butler 1, L S Ramos 1
PMCID: PMC263117  PMID: 8195387

Abstract

Current methods for identifying mycobacteria by high-performance liquid chromatography (HPLC) require a visual assessment of the generated chromatographic data, which often involves time-consuming hand calculations and the use of flow charts. Our laboratory has developed a personal computer-based file containing patterns of mycolic acids detected in 45 species of Mycobacterium, including both slowly and rapidly growing species, as well as Tsukamurella paurometabolum and members of the genera Corynebacterium, Nocardia, Rhodococcus, and Gordona. The library was designed to be used in conjunction with a commercially available pattern recognition software package, Pirouette (Infometrix, Seattle, Wash.). Pirouette uses the K-nearest neighbor algorithm, a similarity-based classification method, to categorize unknown samples on the basis of their multivariate proximities to samples of a preassigned category. Multivariate proximity is calculated from peak height data, while peak heights are named by retention time matching. The system was tested for accuracy by using 24 species of Mycobacterium. Of the 1,333 strains evaluated, > or = 97% were correctly identified. Identification of M. tuberculosis (n = 649) was 99.85% accurate, and identification of the M. avium complex (n = 211) was > or = 98% accurate; > or = 95% of strains of both double-cluster and single-cluster M. gordonae (n = 47) were correctly identified. This system provides a rapid, highly reliable assessment of HPLC-generated chromatographic data for the identification of mycobacteria.

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Selected References

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  1. Butler W. R., Ahearn D. G., Kilburn J. O. High-performance liquid chromatography of mycolic acids as a tool in the identification of Corynebacterium, Nocardia, Rhodococcus, and Mycobacterium species. J Clin Microbiol. 1986 Jan;23(1):182–185. doi: 10.1128/jcm.23.1.182-185.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Butler W. R., Jost K. C., Jr, Kilburn J. O. Identification of mycobacteria by high-performance liquid chromatography. J Clin Microbiol. 1991 Nov;29(11):2468–2472. doi: 10.1128/jcm.29.11.2468-2472.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Butler W. R., Kilburn J. O. High-performance liquid chromatography patterns of mycolic acids as criteria for identification of Mycobacterium chelonae, Mycobacterium fortuitum, and Mycobacterium smegmatis. J Clin Microbiol. 1990 Sep;28(9):2094–2098. doi: 10.1128/jcm.28.9.2094-2098.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Butler W. R., Kilburn J. O. Identification of major slowly growing pathogenic mycobacteria and Mycobacterium gordonae by high-performance liquid chromatography of their mycolic acids. J Clin Microbiol. 1988 Jan;26(1):50–53. doi: 10.1128/jcm.26.1.50-53.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Butler W. R., Kilburn J. O., Kubica G. P. High-performance liquid chromatography analysis of mycolic acids as an aid in laboratory identification of Rhodococcus and Nocardia species. J Clin Microbiol. 1987 Nov;25(11):2126–2131. doi: 10.1128/jcm.25.11.2126-2131.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Butler W. R., O'Connor S. P., Yakrus M. A., Smithwick R. W., Plikaytis B. B., Moss C. W., Floyd M. M., Woodley C. L., Kilburn J. O., Vadney F. S. Mycobacterium celatum sp. nov. Int J Syst Bacteriol. 1993 Jul;43(3):539–548. doi: 10.1099/00207713-43-3-539. [DOI] [PubMed] [Google Scholar]
  7. Butler W. R., Thibert L., Kilburn J. O. Identification of Mycobacterium avium complex strains and some similar species by high-performance liquid chromatography. J Clin Microbiol. 1992 Oct;30(10):2698–2704. doi: 10.1128/jcm.30.10.2698-2704.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Floyd M. M., Silcox V. A., Jones W. D., Jr, Butler W. R., Kilburn J. O. Separation of Mycobacterium bovis BCG from Mycobacterium tuberculosis and Mycobacterium bovis by using high-performance liquid chromatography of mycolic acids. J Clin Microbiol. 1992 May;30(5):1327–1330. doi: 10.1128/jcm.30.5.1327-1330.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Guthertz L. S., Lim S. D., Jang Y., Duffey P. S. Curvilinear-gradient high-performance liquid chromatography for identification of mycobacteria. J Clin Microbiol. 1993 Jul;31(7):1876–1881. doi: 10.1128/jcm.31.7.1876-1881.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Saito H., Tomioka H., Sato K., Tasaka H., Dawson D. J. Identification of various serovar strains of Mycobacterium avium complex by using DNA probes specific for Mycobacterium avium and Mycobacterium intracellulare. J Clin Microbiol. 1990 Aug;28(8):1694–1697. doi: 10.1128/jcm.28.8.1694-1697.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Thibert L., Lapierre S. Routine application of high-performance liquid chromatography for identification of mycobacteria. J Clin Microbiol. 1993 Jul;31(7):1759–1763. doi: 10.1128/jcm.31.7.1759-1763.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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