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Journal of Clinical Microbiology, February 2003, p. 915, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.915.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

LETTER TO THE EDITOR

Outbreak of Infection with Mycobacterium abscessus: Is Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis a Useful Technique for Epidemiological Typing?


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LETTER
 
We have read with interest the article by Zhibang et al. (12) about an outbreak of infections due to Mycobacterium abscessus. We believe, as the authors did, that the strains isolated from the different patients and the environmental samples were the same clone. Classical epidemiology data support this fact, as did conventional microbiology data and plasmid analysis (although only one plasmid was detected and no digestion with restriction enzymes or other studies were done to demonstrate the identity of the plasmids). However, we have some clarifications to make about the use of sodium dodecyl sulfate-polyacyrylamide gel electrophoresis (SDS-PAGE) of whole-cell proteins as an epidemiological marker in this case.

SDS-PAGE of whole-cell proteins is a technique that has been used for epidemiological typing in many cases (1, 3, 9, 11). However, in other cases, the stability of protein profiles made it a technique useful for characterization of bacterial isolates instead of epidemiological typing (5). In the case of rapidly growing mycobacteria, SDS-PAGE of whole-cell proteins has been used by several authors for characterization of the strains, even for taxonomic purposes (2, 6-8). In our experience (4), SDS-PAGE is a useful technique for characterization but not for epidemiology studies. In our study, intraspecies similarity showed Dice coefficients higher than 95%, with only one strain of Mycobacterium fortuitum with a six-band difference. However, interspecies similarity was always below 75%. The same profile could be found between different isolates of the same species from different origins without any relation between them, and identification of the species could be easily performed comparing the profile of the problem strain with the profile of the type strain, when both strains are analyzed in the same gel. Because of all these data, we think that SDS-PAGE is not a useful technique for determining the isogeny of isolates of rapidly growing mycobacteria. Other previously described techniques are more accurate in achieving this objective (10).


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  2. De Jong, A., A. H. Hoentjen, and A. G. Van Der Zanden. 1991. A rapid method for identification of Mycobacterium species by polyacrylamide gel electrophoresis of soluble cell proteins. J. Med. Microbiol. 34:1-5.[Abstract/Free Full Text]
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  3. Esteban, J., I. Gadea, R. Fernandez Roblas, A. Molleja, R. Calvo, V. Acebron, and F. Soriano. 1999. Pseudo-outbreak of Aeromonas hydrophila isolates related to endoscopy. J. Hosp. Infect. 41:313-316.[CrossRef][Medline]
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  4. Esteban, J., A. Molleja, F. Cabria, and M. S. Jimenez. SDS-PAGE for identification of species belonging to the Mycobacterium fortuitum complex. Clin. Microbiol. Infect., in press.
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  5. Esteban, J., E. Nieto, R. Calvo, R. Fernandez Roblas, P. L. Valero Guillen, and F. Soriano. 1999. Microbiological characterization and clinical significance of Corynebacterium amycolatum strains. Eur. J. Clin. Microbiol. Infect. Dis. 18:518-521.[CrossRef][Medline]
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  6. Fevre, A., J. Fougerat, S. Bruneau, and R. Guinet. 1991. Characterization of Mycobacterium species by sodium dodecyl sulfate-polyacrylamide gel elec-trophoresis. Appl. Theor. Electrophor. 2:13-16.[Medline]
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  7. Haas, H., Y. Davidson, and T. Sacks. 1972. Taxonomy of mycobacteria studies by polyacrylamide-gel electrophoresis of cell proteins. J. Med. Microbiol. 5:31-37.[Abstract/Free Full Text]
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  8. Haas, H., J. Michel, and T. Sacks. 1974. Identification of Mycobacterium fortuitum, Mycobacterium abscessus, and Mycobacterium borstelense by polyacrylamide gel electrophoresis of their cell proteins. Int. J. Syst. Bacteriol. 24:366-369.[Abstract/Free Full Text]
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  10. Lai, K. K., B. A. Brown, J. A. Westerling, S. A. Fontecchio, Y. Zhang, and R. J. Wallace, Jr. 1998. Long-term laboratory contamination by Mycobacterium abscessus resulting in two pseudo-outbreaks: recognition with use of random amplified polymorphic DNA (RAPD) polymerase chain reaction. Clin. Infect. Dis. 27:169-175.[Medline]
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  11. McKenzie, H., M. G. Morgan, J. Z. Jordens, M. C. Enright, and M. Bain. 1992. Characterisation of hospital isolates of Moraxella (Branhamella) catarrhalis by SDS-PAGE of whole-cell proteins, immunoblotting and restriction-endonuclease analysis. J. Med. Microbiol. 37:70-76.[Abstract/Free Full Text]
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  12. Zhibang, Y., Z. BiXia, L. Qishan, C. Lihao, L. Xiangquan, and L. Huaping. 2002. Large-scale outbreak of infection with Mycobacterium chelonae subsp. abscessus after penicillin injection. J. Clin. Microbiol. 40:2626-2628.[Abstract/Free Full Text]
J. Esteban*
F. Cabria
Department of Medical Microbiology
Fundación Jiménez Díaz
Av. Reyes Católicos 2
28040-Madrid, Spain

* Phone: 34 915504900Fax: 34 915494764. E-mail: jesteban{at}fjd.es


Author's Reply


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LETTER 
 
I thank Drs. Esteban and Cabria for their interest in our article. We analyzed the isogeny of the isolates mainly depending on patterns of resistance to antibacterial agents and chemical disinfectants, plasmid sizes, and other classical epidemiology data (some not mentioned in our study). Regarding the comments about SDS-PAGE, it has been documented that this is a useful technique for the characterization of species and epidemiological typing. Our study described our first use of SPS-PAGE profiles, and we regard them as helpful but not the only evidence for the isogeny of the isolates, though we lack experience in the use of this technique in epidemiological studies, especially of rapidly growing mycobacteria. I appreciate the comments; our team will do further research related to our epidemiological studies.

Yang Zhibang*
Department of Microbiology Chongqing University of Medical Sciences
Chongqing 400016, China

* Phone: 34 915504900Fax: 34 915494764. E-mail: dryangfm365{at}sina.com

Journal of Clinical Microbiology, February 2003, p. 915, Vol. 41, No. 2
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.2.915.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.




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