1887

Abstract

spp. and other non-dermatophyte fungi are repeatedly isolated from abnormal nails. To investigate whether these fungi are the aetiological agents of infection or simply transient contaminants, a PCR/sequencing/RFLP assay was developed for direct and routine identification of the infecting fungi in onychomycosis. Fungal DNA was readily extracted using a commercial kit after dissolving nail fragments in a NaS solution. Amplification of part of the 28S rDNA by PCR was performed with universal primers and the fungal species were identified by sequencing. The PCR/sequencing results were comparable with microbiological identification from the same nail sample. In addition to dermatophytes, spp. and other less frequently isolated non-dermatophyte fungi were identified as single fungal agents in onychomycosis. Moreover, mixed infections were clearly demonstrated in 10 % of cases by RFLP analysis of PCR products. Identification of infectious agents could be obtained in 2 days, whilst results from fungal cultures take 1–3 weeks. Rapid and reliable molecular identification of the infectious fungus expedites the choice of appropriate antifungal therapy, thereby improving the cure rate of onychomycosis.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.46723-0
2006-09-01
2019-10-23
Loading full text...

Full text loading...

/deliver/fulltext/jmm/55/9/1211.html?itemId=/content/journal/jmm/10.1099/jmm.0.46723-0&mimeType=html&fmt=ahah

References

  1. Baek, S.-C., Chae, H.-J., Houh, D., Byun, D.-G. & Cho, B.-K. ( 1998; ). Detection and differentiation of causative fungi of onychomycosis using PCR amplification and restriction enzyme analysis. Int J Dermatol 37, 682–686.[CrossRef]
    [Google Scholar]
  2. Baudraz-Rosselet, F., Monod, M., Porchet, S. & Frenk, E. ( 1992; ). Retrospective study of the efficacy of two methods of microscopical examination in dermatological mycology. J Mycol Med 2, 148–150.
    [Google Scholar]
  3. Chabasse, D. ( 2004; ). Epidémiologie et étiologie des onychomycoses. In Onychomycoses, pp. 1–35. Edited by R. Baran & G. E. Piérard. Paris: Masson.
  4. de Hoog, G. S., Guarro, J., Gené, J. & Figueras, M. J. ( 2000; ). Atlas of Clinical Fungi, 2nd edn. Utrecht: Centrallbureau voor Schimmelcultures.
  5. English, M. P. ( 1976; ). Nails and fungi. Br J Dermatol 94, 697–701.[CrossRef]
    [Google Scholar]
  6. Garcia-Effron, G., Gomez-Lopez, A., Mellado, E., Monzon, A., Rodriguez-Tudela, J. L. & Cuenca-Estrella, M. ( 2004; ). In vitro activity of terbinafine against medically important non-dermatophyte species of filamentous fungi. J Antimicrob Chemother 53, 1086–1089.[CrossRef]
    [Google Scholar]
  7. Gupta, A. K., Cooper, E. A., MacDonald, P. & Summerbell, R. C. ( 2001a; ). Utility of inoculum counting (Walshe and English criteria) in the clinical diagnosis of onychomycosis caused by nondermatophytic filamentous fungi. J Clin Microbiol 39, 2115–2121.[CrossRef]
    [Google Scholar]
  8. Gupta, A. K., Gregurek-Novak, T., Konnikov, N., Lynde, C. W., Hofstader, S. & Summerbell, R. C. ( 2001b; ). Itraconazole and terbinafine treatment of some nondermatophyte molds causing onychomycosis of the toes and a review of the literature. J Cutan Med Surg 5, 206–210.[CrossRef]
    [Google Scholar]
  9. Kane, J., Scott, J. A., Summerbell, R. C. & Diena, B. ( 1992; ). Trichophyton krajdenii sp. nov. an anthropophilic dermatophyte. Mycotaxon 45, 307–316.
    [Google Scholar]
  10. Kardjeva, V., Summerbell, R., Kantardjiev, T., Devliotou-Panagiotidou, D., Sotiriou, E. & Graser, Y. ( 2006; ). Forty-eight-hour diagnosis of onychomycosis with subtyping of Trichophyton rubrum strains. J Clin Microbiol 44, 1419–1427.[CrossRef]
    [Google Scholar]
  11. Machouart-Dubach, M., Lacroix, C., Feuilhade de Chauvin, M., Le Gall, I., Giudicelli, C., Lorenzo, F. & Derouin, F. ( 2001; ). Rapid discrimination among dermatophytes, Scytalidium spp., and other fungi with a PCR-restriction fragment length polymorphism ribotyping method. J Clin Microbiol 39, 685–690.[CrossRef]
    [Google Scholar]
  12. Menotti, J., Machouart, M., Benderdouche, M., Cetre-Sossah, C., Morel, P., Dubertret, L., Derouin, F., Feuilhade De Chauvin, M. & Lacroix, C. ( 2004; ). Polymerase chain reaction for diagnosis of dermatophyte and Scytalidium spp. onychomycosis. Br J Dermatol 151, 518–519.[CrossRef]
    [Google Scholar]
  13. Mock, M., Monod, M., Baudraz-Rosselet, F. & Panizzon, R. G. ( 1998; ). Tinea capitis dermatophytes: susceptibility to antifungal drugs tested in vitro and in vivo. Dermatology 197, 361–367.[CrossRef]
    [Google Scholar]
  14. Monod, M., Baudraz-Rosselet, F., Ramelet, A. A. & Frenk, E. ( 1989; ). Direct mycological examination in dermatology: a comparison of different methods. Dermatologica 179, 183–186.[CrossRef]
    [Google Scholar]
  15. Ninet, B., Jan, I., Bontems, O., Léchenne, B., Jousson, O., Panizzon, R., Lew, D. & Monod, M. ( 2003; ). Identification of dermatophyte species by 28S ribosomal DNA sequencing with a commercial kit. J Clin Microbiol 41, 826–830.[CrossRef]
    [Google Scholar]
  16. Scherer, W. P., McCreary, J. P. & Hayes, W. W. ( 2001; ). The diagnosis of onychomycosis in a geriatric population: a study of 450 cases in South Florida. J Am Podiatr Med Assoc 91, 456–464.[CrossRef]
    [Google Scholar]
  17. Summerbell, R. C., Kane, J. & Kradjen, S. ( 1989; ). Onychomycosis, tinea pedis and tinea manuum caused by non-dermatophytic filamentous fungi. Mycoses 32, 609–619.
    [Google Scholar]
  18. Summerbell, R. C., Haugland, R. A., Li, A. & Gupta, A. K. ( 1999; ). rRNA gene internal transcribed spacer 1 and 2 sequences of asexual, anthropophilic dermatophytes related to Trichophyton rubrum. J Clin Microbiol 37, 4005–4011.
    [Google Scholar]
  19. Summerbell, R. C., Cooper, E., Bunn, U., Jamieson, F. & Gupta, A. K. ( 2005; ). Onychomycosis: a critical study of techniques and criteria for confirming the etiologic significance of nondermatophytes. Med Mycol 43, 39–59.[CrossRef]
    [Google Scholar]
  20. Tosti, A., Piraccini, B. M. & Lorenzi, S. ( 2000; ). Onychomycosis caused by nondermatophytic molds: clinical features and response to treatment of 59 cases. J Am Acad Dermatol 42, 217–224.[CrossRef]
    [Google Scholar]
  21. Turin, L., Riva, F., Galbiati, G. & Cainelli, T. ( 2000; ). Fast, simple and highly sensitive double-rounded polymerase chain reaction assay to detect medically relevant fungi in dermatological specimens. Eur J Clin Invest 30, 511–518.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.46723-0
Loading
/content/journal/jmm/10.1099/jmm.0.46723-0
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