Abstract
Nineteen reference and 156 clinical strains of the genus Nocardia belonging to 12 taxonomic groups were studied for restriction fragment length polymorphism (RFLP) by using an amplified 439-bp segment of the 65-kDa heat shock protein gene. Of 30 restriction endonucleases, digestion with MspI and then digestion with BsaHI produced RFLP band patterns which separated all 12 groups except N. asteroides type IV from 6 of 12 N. transvalensis isolates and N. carnea from the N. asteroides type VI isolates. Commonly encountered species such as N. nova, N. farcinica, N. brasiliensis sensu stricto, and N. otitidiscaviarum were easily separated. Each taxon resulted in a single RFLP band pattern that included > or = 96% of all biochemically grouped isolates for 9 of 12 taxa with MspI and for 8 of 12 taxa with BsaHI. With the use of both patterns, only 6 of 175 (3.4%) isolates failed to fit the biochemically defined group patterns. These studies provide the first evidence for the separate identities of four antibiogram-defined (but currently unnamed) groups within the N. asteroides complex (types I, II, IV, and VI) and the presence of two subgroups within N. transvalensis. They also provide genotypic evidence for the separate identities of N. nova and N. farcinica. The lack of BstEII recognition sites in amplicons obtained from nocardiae provides a simple and rapid method for the differentiation of nocardiae from mycobacteria. DNA amplification with RFLP analysis is the first rapid method that distinguishes all clinically significant taxa and recognized species within the genus Nocardia.
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- 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]
- 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]
- 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]
- Chun J., Goodfellow M. A phylogenetic analysis of the genus Nocardia with 16S rRNA gene sequences. Int J Syst Bacteriol. 1995 Apr;45(2):240–245. doi: 10.1099/00207713-45-2-240. [DOI] [PubMed] [Google Scholar]
- Cook S. M., Bartos R. E., Pierson C. L., Frank T. S. Detection and characterization of atypical mycobacteria by the polymerase chain reaction. Diagn Mol Pathol. 1994 Mar;3(1):53–58. doi: 10.1097/00019606-199403010-00009. [DOI] [PubMed] [Google Scholar]
- Inyaku K., Hiyama K., Ishioka S., Inamizu T., Yamakido M. Rapid detection and identification of mycobacteria in sputum samples by nested polymerase chain reaction and restriction fragment length polymorphisms of dnaJ heat shock protein gene. Hiroshima J Med Sci. 1993 Mar;42(1):21–31. [PubMed] [Google Scholar]
- Kirschner P., Kiekenbeck M., Meissner D., Wolters J., Böttger E. C. Genetic heterogeneity within Mycobacterium fortuitum complex species: genotypic criteria for identification. J Clin Microbiol. 1992 Nov;30(11):2772–2775. doi: 10.1128/jcm.30.11.2772-2775.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lungu O., Della Latta P., Weitzman I., Silverstein S. Differentiation of Nocardia from rapidly growing Mycobacterium species by PCR-RFLP analysis. Diagn Microbiol Infect Dis. 1994 Jan;18(1):13–18. doi: 10.1016/0732-8893(94)90128-7. [DOI] [PubMed] [Google Scholar]
- McNeil M. M., Brown J. M., Georghiou P. R., Allworth A. M., Blacklock Z. M. Infections due to Nocardia transvalensis: clinical spectrum and antimicrobial therapy. Clin Infect Dis. 1992 Sep;15(3):453–463. doi: 10.1093/clind/15.3.453. [DOI] [PubMed] [Google Scholar]
- McNeil M. M., Brown J. M., Magruder C. H., Shearlock K. T., Saul R. A., Allred D. P., Ajello L. Disseminated Nocardia transvalensis infection: an unusual opportunistic pathogen in severely immunocompromised patients. J Infect Dis. 1992 Jan;165(1):175–178. doi: 10.1093/infdis/165.1.175. [DOI] [PubMed] [Google Scholar]
- Mishra S. K., Gordon R. E., Barnett D. A. Identification of nocardiae and streptomycetes of medical importance. J Clin Microbiol. 1980 Jun;11(6):728–736. doi: 10.1128/jcm.11.6.728-736.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Plikaytis B. B., Plikaytis B. D., Yakrus M. A., Butler W. R., Woodley C. L., Silcox V. A., Shinnick T. M. Differentiation of slowly growing Mycobacterium species, including Mycobacterium tuberculosis, by gene amplification and restriction fragment length polymorphism analysis. J Clin Microbiol. 1992 Jul;30(7):1815–1822. doi: 10.1128/jcm.30.7.1815-1822.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Soini H., Böttger E. C., Viljanen M. K. Identification of mycobacteria by PCR-based sequence determination of the 32-kilodalton protein gene. J Clin Microbiol. 1994 Dec;32(12):2944–2947. doi: 10.1128/jcm.32.12.2944-2947.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Springer B., Böttger E. C., Kirschner P., Wallace R. J., Jr Phylogeny of the Mycobacterium chelonae-like organism based on partial sequencing of the 16S rRNA gene and proposal of Mycobacterium mucogenicum sp. nov. Int J Syst Bacteriol. 1995 Apr;45(2):262–267. doi: 10.1099/00207713-45-2-262. [DOI] [PubMed] [Google Scholar]
- Steingrube V. A., Gibson J. L., Brown B. A., Zhang Y., Wilson R. W., Rajagopalan M., Wallace R. J., Jr PCR amplification and restriction endonuclease analysis of a 65-kilodalton heat shock protein gene sequence for taxonomic separation of rapidly growing mycobacteria. J Clin Microbiol. 1995 Jan;33(1):149–153. doi: 10.1128/jcm.33.1.149-153.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steingrube V. A., Wallace R. J., Jr, Brown B. A., Pang Y., Zeluff B., Steele L. C., Zhang Y. Acquired resistance of Nocardia brasiliensis to clavulanic acid related to a change in beta-lactamase following therapy with amoxicillin-clavulanic acid. Antimicrob Agents Chemother. 1991 Mar;35(3):524–528. doi: 10.1128/aac.35.3.524. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Telenti A., Marchesi F., Balz M., Bally F., Böttger E. C., Bodmer T. Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis. J Clin Microbiol. 1993 Feb;31(2):175–178. doi: 10.1128/jcm.31.2.175-178.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Vaneechoutte M., De Beenhouwer H., Claeys G., Verschraegen G., De Rouck A., Paepe N., Elaichouni A., Portaels F. Identification of Mycobacterium species by using amplified ribosomal DNA restriction analysis. J Clin Microbiol. 1993 Aug;31(8):2061–2065. doi: 10.1128/jcm.31.8.2061-2065.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wallace R. J., Jr, Brown B. A., Blacklock Z., Ulrich R., Jost K., Brown J. M., McNeil M. M., Onyi G., Steingrube V. A., Gibson J. New Nocardia taxon among isolates of Nocardia brasiliensis associated with invasive disease. J Clin Microbiol. 1995 Jun;33(6):1528–1533. doi: 10.1128/jcm.33.6.1528-1533.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wallace R. J., Jr, Brown B. A., Tsukamura M., Brown J. M., Onyi G. O. Clinical and laboratory features of Nocardia nova. J Clin Microbiol. 1991 Nov;29(11):2407–2411. doi: 10.1128/jcm.29.11.2407-2411.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wallace R. J., Jr, Steele L. C., Sumter G., Smith J. M. Antimicrobial susceptibility patterns of Nocardia asteroides. Antimicrob Agents Chemother. 1988 Dec;32(12):1776–1779. doi: 10.1128/aac.32.12.1776. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wallace R. J., Jr, Steele L. C. Susceptibility testing of Nocardia species for the clinical laboratory. Diagn Microbiol Infect Dis. 1988 Mar;9(3):155–166. doi: 10.1016/0732-8893(88)90025-9. [DOI] [PubMed] [Google Scholar]
- Wallace R. J., Jr, Tsukamura M., Brown B. A., Brown J., Steingrube V. A., Zhang Y. S., Nash D. R. Cefotaxime-resistant Nocardia asteroides strains are isolates of the controversial species Nocardia farcinica. J Clin Microbiol. 1990 Dec;28(12):2726–2732. doi: 10.1128/jcm.28.12.2726-2732.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]