This Article
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 Devallois, A.
Right arrow Articles by Rastogi, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Devallois, A.
Right arrow Articles by Rastogi, N.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, 11 1997, 2767-2772, Vol 35, No. 11
Copyright © 1997 by the American Society for Microbiology. All rights reserved.

Molecular characterization of Mycobacterium avium complex isolates giving discordant results in AccuProbe tests by PCR-restriction enzyme analysis, 16S rRNA gene sequencing, and DT1-DT6 PCR [In Process Citation]

A Devallois, M Picardeau, CN Paramasivan, V Vincent and N Rastogi
Unite de la Tuberculose et des Mycobacteries, Institut Pasteur, Pointe- a-Pitre, Guadeloupe, French West Indies.

Based on cultural and biochemical tests, a total of 84 strains (72 clinical and 12 environmental isolates from the Caribbean Isles, Europe, and the Indian subcontinent) were identified as members of the Mycobacterium avium complex (MAC). They were further characterized with MAC, M. avium, and M. intracellulare probes of the AccuProbe system, and this was followed by selective amplification of DT6 and DT1 sequences. Seventy isolates gave concordant results; 63 were identified as M. avium, 5 were identified as M. intracellulare, and 24 remained untypeable by both methods. Fourteen isolates gave discrepant results, as they were DT1 positive but gave negative results by the M. intracellulare AccuProbe test. Consequently, a detailed molecular analysis of all DT1-positive isolates (14 discrepant strains plus 5 M. intracellulare strains) was performed by PCR-restriction analysis (PRA) of the hsp65 gene and 16S rRNA gene sequencing. The results confirmed the reported heterogeneity of M. intracellulare, as only 6 of 19 isolates (32%) gave PRA results compatible with published M. intracellulare profiles while the rest of the isolates were grouped in four previously unpublished profiles. 16S rRNA gene sequencing showed that only 8 of 19 isolates (42%) were related to M. intracellulare IWGMT 90247 (EMBL accession no. X88917), the rest being related to MCRO19 (EMBL accession no. X93030) and MIWGTMR10 (EMBL accession no. X88915). In conclusion, we have characterized a significant number of MAC isolates which were not identified by the AccuProbe test, PRA, or 16S rRNA sequencing. However, all of them were identifiable by DT1-DT6 PCR (they were DT6 negative and DT1 positive) and could be tentatively identified as M. intracellulare based on previously published observations. It is noteworthy that the majority of such isolates (14 of 19) were from the Indian subcontinent, with 12 of 14 being environmental isolates. Our study confirms the marked heterogeneity of M. intracellulare isolates and shows the utility of in-house DT1 PCR to detect this group of isolates, which would otherwise have been missed by the AccuProbe system in a routine clinical microbiology laboratory.


This article has been cited by other articles:

  • Richardson, E. T., Samson, D., Banaei, N. (2009). Rapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR. J. Clin. Microbiol. 47: 1497-1502 [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]  
  • Prammananan, T., Phunpruch, S., Tingtoy, N., Srimuang, S., Chaiprasert, A. (2006). Distribution of hsp65 PCR-Restriction Enzyme Analysis Patterns among Mycobacterium avium Complex Isolates in Thailand. J. Clin. Microbiol. 44: 3819-3821 [Abstract] [Full Text]  
  • Huang, T.-S., Lee, S. S.-J., Chen, Y.-S., Tu, H.-Z., Huang, W.-K., Liu, Y.-C. (2005). Discordant molecular characterization results in a Mycobacterium avium complex strain isolated from an AIDS patient. J Med Microbiol 54: 681-683 [Abstract] [Full Text]  
  • Lebrun, L., Weill, F.-X., Lafendi, L., Houriez, F., Casanova, F., Gutierrez, M. C., Ingrand, D., Lagrange, P., Vincent, V., Herrmann, J. L. (2005). Use of the INNO-LiPA-MYCOBACTERIA Assay (Version 2) for Identification of Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum Complex Isolates. J. Clin. Microbiol. 43: 2567-2574 [Abstract] [Full Text]  
  • Adekambi, T., Colson, P., Drancourt, M. (2003). rpoB-Based Identification of Nonpigmented and Late-Pigmenting Rapidly Growing Mycobacteria. J. Clin. Microbiol. 41: 5699-5708 [Abstract] [Full Text]  
  • Vanitha, J. D., Venkatasubramani, R., Dharmalingam, K., Paramasivan, C. N. (2003). Large-Restriction-Fragment Polymorphism Analysis of Mycobacterium chelonae and Mycobacterium terrae Isolates. Appl. Environ. Microbiol. 69: 4337-4341 [Abstract] [Full Text]  
  • da Silva Rocha, A., Werneck Barreto, A. M., Dias Campos, C. E., Villas-Boas da Silva, M., Fonseca, L., Saad, M. H., Degrave, W. M., Suffys, P. N. (2002). Novel Allelic Variants of Mycobacteria Isolated in Brazil as Determined by PCR-Restriction Enzyme Analysis of hsp65. J. Clin. Microbiol. 40: 4191-4196 [Abstract] [Full Text]  
  • Smole, S. C., McAleese, F., Ngampasutadol, J., von Reyn, C. F., Arbeit, R. D. (2002). Clinical and Epidemiological Correlates of Genotypes within the Mycobacterium avium Complex Defined by Restriction and Sequence Analysis of hsp65. J. Clin. Microbiol. 40: 3374-3380 [Abstract] [Full Text]  
  • Menendez, M. C., Palenque, E., Navarro, M. C., Nunez, M. C., Rebollo, M. J., Garcia, M. J. (2001). Characterization of a Mycobacterium intracellulare Variant Strain by Molecular Techniques. J. Clin. Microbiol. 39: 4241-4246 [Abstract] [Full Text]  
  • Suffys, P. N., da Silva Rocha, A., de Oliveira, M., Dias Campos, C. E., Werneck Barreto, A. M., Portaels, F., Rigouts, L., Wouters, G., Jannes, G., van Reybroeck, G., Mijs, W., Vanderborght, B. (2001). Rapid Identification of Mycobacteria to the Species Level Using INNO-LiPA Mycobacteria, a Reverse Hybridization Assay. J. Clin. Microbiol. 39: 4477-4482 [Abstract] [Full Text]  
  • Kim, B.-J., Lee, K.-H., Park, B.-N., Kim, S.-J., Bai, G.-H., Kim, S.-J., Kook, Y.-H. (2001). Differentiation of Mycobacterial Species by PCR-Restriction Analysis of DNA (342 Base Pairs) of the RNA Polymerase Gene (rpoB). J. Clin. Microbiol. 39: 2102-2109 [Abstract] [Full Text]  
  • Oliveira, R. S., Sircili, M. P., Ueki, S. Y. M., Telles, M. A. S., Schnabel, B., Briones, M. R. S., Leão, S. C. (2000). PCR-Restriction Enzyme Analysis of a Bone Marrow Isolate from a Human Immunodeficiency Virus-Positive Patient Discloses Polyclonal Infection with Two Mycobacterium avium Strains. J. Clin. Microbiol. 38: 4643-4645 [Abstract] [Full Text]  
  • Leão, S. C., Briones, M. R. S., Sircili, M. P., Balian, S. C., Mores, N., Ferreira-Neto, J. S. (1999). Identification of Two Novel Mycobacterium avium Allelic Variants in Pig and Human Isolates from Brazil by PCR-Restriction Enzyme Analysis. J. Clin. Microbiol. 37: 2592-2597 [Abstract] [Full Text]  
  • Kim, B.-J., Lee, S.-H., Lyu, M.-A., Kim, S.-J., Bai, G.-H., Kim, S.-J., Chae, G.-T., Kim, E.-C., Cha, C.-Y., Kook, Y.-H. (1999). Identification of Mycobacterial Species by Comparative Sequence Analysis of the RNA Polymerase Gene (rpoB). J. Clin. Microbiol. 37: 1714-1720 [Abstract] [Full Text]