1887

Abstract

In order to identify the source of infections caused by dermatophytes, as well as the pathogen transmission pathway, there is a need to determine methods that allow detailed genetic differentiation of the strains within the dermatophyte genera. In this work, a PCR melting profile (PCR-MP) technique based on the ligation of adaptors and the difference in melting temperatures of DNA restriction fragments was used for the first time for intraspecies genotyping of dermatophytes. Clinical isolates and reference strains of dermatophytes isolated from skin, scalp, toenails and fingernails were used for this study. PCR-MP and random amplification of polymorphic DNA (RAPD) were used to type 11 isolates of , 40 isolates of and 14 isolates of . The results distinguished five types (containing one subtype) characteristic for and seven types characteristic for using the PCR-MP technique. Analysis conducted using RAPD revealed five types for and four types for isolates. No differentiation was observed for the isolates with either method. These results demonstrate that PCR-MP is a reliable method for the differentiation of and strains and yields a discriminatory power that is at least equal to that of RAPD.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.013458-0
2010-02-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jmm/59/2/185.html?itemId=/content/journal/jmm/10.1099/jmm.0.013458-0&mimeType=html&fmt=ahah

References

  1. Abdel-Rahman S. M. 2008; Strain differentiation of dermatophytes. Mycopathologia 166:319–333 [CrossRef]
    [Google Scholar]
  2. Baeza L. C., Giannini M. J. 2004; Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD. Rev Inst Med Trop Sao Paulo 46:339–341 [CrossRef]
    [Google Scholar]
  3. Baeza L. C., Matsumoto M. T., Almeida A. M. F., Mendes-Gianini M. J. S. 2006; Strain differentiation of Trichophyton rubrum by randomly amplified polymorphic DNA and analysis of rDNA nontranscribed spacer. J Med Microbiol 55:429–436 [CrossRef]
    [Google Scholar]
  4. Brilhante R. S., Cordeiro R. A., Medrano D. J., Monteiro A. J., Sidrim J. J., Rocha M. F. 2005; Antifungal susceptibility and genotypical pattern of Microsporum canis strains. Can J Microbiol 51:507–510 [CrossRef]
    [Google Scholar]
  5. de Assis Santos D., de Carvalho Araújo R. A., Kohler L. M., Machado-Pinto J., Hamdan J. S., Cisalpino P. S. 2007; Molecular typing and antifungal susceptibility of Trichophyton rubrum isolates from patients with onychomycosis pre- and post-treatment. Int J Antimicrob Agents 29:563–569 [CrossRef]
    [Google Scholar]
  6. Dobrowolska A., Staczek P., Kaszuba A., Kozłowska M. 2006; PCR-RFLP analysis of the dermatophytes isolated from patients in Central Poland. J Dermatol Sci 42:71–74 [CrossRef]
    [Google Scholar]
  7. Ellsworth D. L., Rittenhouse K. D., Honeycutt R. L. 1993; Artifactual variation in randomly amplified polymorphic DNA banding patterns. Biotechniques 14:214–217
    [Google Scholar]
  8. Faggi E., Pini G., Campisi E., Bertellini C., Difonzo E., Mancianti F. 2001; Application of PCR to distinguish common species of dermatophytes. J Clin Microbiol 39:3382–3385 [CrossRef]
    [Google Scholar]
  9. Gräser Y., Kuijpers A. F. A., Presber W., Hoog G. S. 2000; Molecular taxonomy of the Trichophyton rubrum complex. J Clin Microbiol 38:3329–3336
    [Google Scholar]
  10. Jackson C. J., Barton R. C., Kelly S. L., Evans E. 2000; Strain identification of Trichophyton rubrum by specific amplification of subrepeat elements in the ribosomal DNA nontranscribed spacer. J Clin Microbiol 38:4527–4534
    [Google Scholar]
  11. Kac G., Bougnoux M. E., Feulhade De Chauvin M., Sene S., Derouin F. 1999; Genetic diversity among Trichophyton mentagrophytes isolates using random amplified polymorphic DNA method. Br J Dermatol 140:839–844 [CrossRef]
    [Google Scholar]
  12. Kamiya A., Kikuchi A., Tomita Y., Kanbe T. 2004; PCR and PCR-RFLP techniques targeting the DNA topoisomerase II gene for rapid clinical diagnosis of the etiologic agent of dermatophytosis. J Dermatol Sci 34:35–38 [CrossRef]
    [Google Scholar]
  13. Kaszubiak A., Klein S., de Hoog G. S., Gräser Y. 2004; Population structure and evolutionary origins of Microsporum canis , M. ferrugineum and M. audouinii . Infect Genet Evol 4:179–186 [CrossRef]
    [Google Scholar]
  14. Krawczyk B, Samet A, Leibner J, Śledzińska A, Kur J. 2006; Evaluation of a PCR melting profile technique for bacterial strain differentiation. J Clin Microbiol 44:2327–2332 [CrossRef]
    [Google Scholar]
  15. Krawczyk B., Leibner J., Stojowska K., Bronk M., Samet A., Kur J. 2007a; PCR melting profile method for genotyping of vancomycin-resistant Enterococcus faecium isolates from hematological unit patients. Pol J Microbiol 56:65–70
    [Google Scholar]
  16. Krawczyk B., Leibner J., Barańska-Rybak W., Samet A., Nowicki R., Kur J. 2007b; ADSRRS-fingerprinting and PCR MP techniques for studies of intraspecies genetic relatedness in Staphylococcus aureus . J Microbiol Methods 71:114–122 [CrossRef]
    [Google Scholar]
  17. Liu D., Coloe S., Baird R., Pederson J. 2000; Rapid mini-preparation of fungal DNA for PCR. J Clin Microbiol 38:471
    [Google Scholar]
  18. Macura A. B. 2006; Nail susceptibility to ex vivo experimental fungal infections. Mikologia Lekarska 13:255–260
    [Google Scholar]
  19. Masny A., Płucienniczak A. 2003; Ligation mediated PCR performed at low denaturation temperatures – PCR melting profiles. Nucleic Acids Res 31:e114 [CrossRef]
    [Google Scholar]
  20. Mochizuki T., Ishizaki H., Barton R. C., Moore M. K., Jackson C. J., Kelly S. L., Evans E. G. V. 2003; Restriction fragment length polymorphism analysis of ribosomal DNA intergenic regions is useful for differentiating strains of Trichophyton mentagrophytes . J Clin Microbiol 41:4583–4588 [CrossRef]
    [Google Scholar]
  21. Sharma R., de Hoog S., Presber W., Gräser Y. 2007; A virulent genotype of Microsporum canis is responsible for the majority of human infections. J Med Microbiol 56:1377–1385 [CrossRef]
    [Google Scholar]
  22. Weitzman I., Summerbell R. 1995; The dermatophytes. Clin Microbiol Rev 8:240–259
    [Google Scholar]
  23. Yazdanparast A., Jackson C. J., Barton R. C., Evans E. G. V. 2003; Molecular strain typing of Trichophyton rubrum indicates multiple strain involvement in onychomycosis. Br J Dermatol 148:51–54 [CrossRef]
    [Google Scholar]
  24. Yu J., Wan Z., Chen W., Wang W., Li R. 2004; Molecular typing study of the Microsporum canis strains isolated from an outbreak of tinea capitis in a school. Mycopathologia 157:37–41 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.013458-0
Loading
/content/journal/jmm/10.1099/jmm.0.013458-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error