This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by McNabb, A.
Right arrow Articles by Isaac-Renton, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by McNabb, A.
Right arrow Articles by Isaac-Renton, J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 2004, p. 3000-3011, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3000-3011.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Assessment of Partial Sequencing of the 65-Kilodalton Heat Shock Protein Gene (hsp65) for Routine Identification of Mycobacterium Species Isolated from Clinical Sources

Alan McNabb,1* Diane Eisler,1 Kathy Adie,1 Marie Amos,1 Mabel Rodrigues,1 Gwen Stephens,1,2 William A. Black,1,2 and Judith Isaac-Renton1,2

Laboratory Services, British Columbia Centre for Disease Control,1 Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada2

Received 14 November 2003/ Returned for modification 27 January 2004/ Accepted 4 April 2004

We assessed the ability of an in-house database, consisting of 111 hsp65 sequences from putative and valid Mycobacterium species or described groups, to identify 689 mycobacterial clinical isolates from 35 species or groups. A preliminary assessment indicated that hsp65 sequencing confirmed the identification of 79.4% of the isolates from the 32 species examined, including all Mycobacterium tuberculosis complex isolates, all isolates from 13 other species, and 95.6% of all M. avium-M. intracellulare complex isolates. Identification discrepancies were most frequently encountered with isolates submitted as M. chelonae, M. fortuitum, M. gordonae, M. scrofulaceum, and M. terrae. Reexamination of isolates with discrepant identifications confirmed that hsp65 identifications were correct in a further 40 isolates. This brought the overall agreement between hsp65 sequencing and the other identification methods to 85.2%. The remaining 102 isolates had sequence matches below our acceptance criterion, had nondifferential sequence matches between two or more species, were identified by 16S rRNA sequencing as a putative taxonomic group not contained in our database, or were identified by hsp65 and 16S rRNA gene sequencing as a species not in our biochemical test database or had conflicting identifications. Therefore, to incorporate the unconfirmed isolates it was necessary to create 29 additional entries in our hsp65 identification database: 18 associated with valid species, 7 indicating unique sequences not associated with valid or putative species or groups, and 4 associated with unique, but currently described taxonomic groups. Confidence in the hsp65 sequence identification of a clinical isolate is best when sequence matches of 100% occur, but our data indicate that correct identifications can be confidently made when unambiguous matches exceeding 97% occur, but are dependent on the completeness of the database. Our study indicates that for hsp65 sequencing to be an effective means for identifying mycobacteria a comprehensive database must be constructed. hsp65 sequencing has the advantage of being more rapid and less expensive than biochemical test panels, uses a single set of reagents to identify both rapid- and slow-growing mycobacteria, and can provide a more definitive identification.

* Corresponding author. Mailing address: Molecular Services, British Columbia Centre for Disease Control, 655 West 12th Ave., Vancouver, British Columbia V5Z 4R4, Canada. Phone: (604) 660-0876. Fax: (604) 660-6073. E-mail: alan.mcnabb{at}bccdc.ca.

Journal of Clinical Microbiology, July 2004, p. 3000-3011, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3000-3011.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Leao, S. C., Tortoli, E., Viana-Niero, C., Ueki, S. Y. M., Lima, K. V. B., Lopes, M. L., Yubero, J., Menendez, M. C., Garcia, M. J. (2009). Characterization of Mycobacteria from a Major Brazilian Outbreak Suggests that Revision of the Taxonomic Status of Members of the Mycobacterium chelonae-M. abscessus Group Is Needed. J. Clin. Microbiol. 47: 2691-2698 [Abstract] [Full Text]  
  • Duarte, R. S., Lourenco, M. C. S., Fonseca, L. d. S., Leao, S. C., Amorim, E. d. L. T., Rocha, I. L. L., Coelho, F. S., Viana-Niero, C., Gomes, K. M., da Silva, M. G., de Oliveira Lorena, N. S., Pitombo, M. B., Ferreira, R. M. C., de Oliveira Garcia, M. H., de Oliveira, G. P., Lupi, O., Vilaca, B. R., Serradas, L. R., Chebabo, A., Marques, E. A., Teixeira, L. M., Dalcolmo, M., Senna, S. G., Sampaio, J. L. M. (2009). Epidemic of Postsurgical Infections Caused by Mycobacterium massiliense. J. Clin. Microbiol. 47: 2149-2155 [Abstract] [Full Text]  
  • Tortoli, E., Baruzzo, S., Heijdra, Y., Klenk, H.-P., Lauria, S., Mariottini, A., van Ingen, J. (2009). Mycobacterium insubricum sp. nov.. Int. J. Syst. Evol. Microbiol. 59: 1518-1523 [Abstract] [Full Text]  
  • Tortoli, E., Adriani, B., Baruzzo, S., Degl'Innocenti, R., Galanti, I., Lauria, S., Mariottini, A., Pascarella, M. (2009). Pulmonary Disease Due to Mycobacterium arosiense, an Easily Misidentified Pathogenic Novel Mycobacterium. J. Clin. Microbiol. 47: 1947-1949 [Abstract] [Full Text]  
  • Leader, B. T., Frye, J. G., Hu, J., Fedorka-Cray, P. J., Boyle, D. S. (2009). High-Throughput Molecular Determination of Salmonella enterica Serovars by Use of Multiplex PCR and Capillary Electrophoresis Analysis. J. Clin. Microbiol. 47: 1290-1299 [Abstract] [Full Text]  
  • Bang, D., Herlin, T., Stegger, M., Andersen, A. B., Torkko, P., Tortoli, E., Thomsen, V. O. (2008). Mycobacterium arosiense sp. nov., a slowly growing, scotochromogenic species causing osteomyelitis in an immunocompromised child. Int. J. Syst. Evol. Microbiol. 58: 2398-2402 [Abstract] [Full Text]  
  • Lazzarini, L. C. O., Huard, R. C., Boechat, N. L., Gomes, H. M., Oelemann, M. C., Kurepina, N., Shashkina, E., Mello, F. C. Q., Gibson, A. L., Virginio, M. J., Marsico, A. G., Butler, W. R., Kreiswirth, B. N., Suffys, P. N., Lapa e Silva, J. R., Ho, J. L. (2007). Discovery of a Novel Mycobacterium tuberculosis Lineage That Is a Major Cause of Tuberculosis in Rio de Janeiro, Brazil. J. Clin. Microbiol. 45: 3891-3902 [Abstract] [Full Text]  
  • Jang, E.-Y., Lee, S.-O., Choi, S.-H., Sung, H., Kim, M.-N., Kim, B.-J., Choi, S.-H., Kim, Y. S., Woo, J. H. (2007). Case of Pyomyositis Due to Mycobacterium haemophilum in a Renal Transplant Recipient. J. Clin. Microbiol. 45: 3847-3849 [Abstract] [Full Text]  
  • Mignard, S., Flandrois, J.-P. (2007). Identification of Mycobacterium using the EF-Tu encoding (tuf) gene and the tmRNA encoding (ssrA) gene. J Med Microbiol 56: 1033-1041 [Abstract] [Full Text]  
  • Turenne, C. Y., Wallace, R. Jr., Behr, M. A. (2007). Mycobacterium avium in the Postgenomic Era. Clin. Microbiol. Rev. 20: 205-229 [Abstract] [Full Text]  
  • Griffith, D. E., Aksamit, T., Brown-Elliott, B. A., Catanzaro, A., Daley, C., Gordin, F., Holland, S. M., Horsburgh, R., Huitt, G., Iademarco, M. F., Iseman, M., Olivier, K., Ruoss, S., von Reyn, C. F., Wallace, R. J. Jr., Winthrop, K., on behalf of the ATS Mycobacterial Diseases Subcom, (2007). An Official ATS/IDSA Statement: Diagnosis, Treatment, and Prevention of Nontuberculous Mycobacterial Diseases. Am. J. Respir. Crit. Care Med. 175: 367-416 [Full Text]  
  • Reischl, U., Melzl, H., Kroppenstedt, R. M., Miethke, T., Naumann, L., Mariottini, A., Mazzarelli, G., Tortoli, E. (2006). Mycobacterium monacense sp. nov.. Int. J. Syst. Evol. Microbiol. 56: 2575-2578 [Abstract] [Full Text]  
  • Sampaio, J. L. M., Junior, D. N., de Freitas, D., Hofling-Lima, A. L., Miyashiro, K., Alberto, F. L., Leao, S. C. (2006). An Outbreak of Keratitis Caused by Mycobacterium immunogenum.. J. Clin. Microbiol. 44: 3201-3207 [Abstract] [Full Text]  
  • Turenne, C. Y., Semret, M., Cousins, D. V., Collins, D. M., Behr, M. A. (2006). Sequencing of hsp65 Distinguishes among Subsets of the Mycobacterium avium Complex. J. Clin. Microbiol. 44: 433-440 [Abstract] [Full Text]  
  • McNabb, A., Adie, K., Rodrigues, M., Black, W. A., Isaac-Renton, J. (2006). Direct Identification of Mycobacteria in Primary Liquid Detection Media by Partial Sequencing of the 65-Kilodalton Heat Shock Protein Gene. J. Clin. Microbiol. 44: 60-66 [Abstract] [Full Text]  
  • Zelazny, A. M., Calhoun, L. B., Li, L., Shea, Y. R., Fischer, S. H. (2005). Identification of Mycobacterium Species by secA1 Sequences. J. Clin. Microbiol. 43: 1051-1058 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»