1887

Abstract

Tinea unguium, known as onychomycosis, is a dermatophyte infection of nails with worldwide distribution. Conventional methods for detecting fungi in nail specimens are either non-specific (microscopy) or insensitive (culture). PCR has been used to improve sensitivity in detecting the causative fungi in nail specimens from patients with suspected onychomycosis. Results of a commercial multiplex PCR for the detection of dermatophytes, especially (the main dermatophyte implicated), as compared to conventional methods are presented. A total of 418 nail scrapings obtained from dermatological outpatients were handled in the Laboratory of Microbiology between May 2010 and May 2013. Among them, multiplex PCR detected 126 (30.1 %) dermatophyte-positive samples, whereas culture revealed 44 (10.5 %). Direct microscopy revealed 63 (15.1 %) positive specimens. was identified in 116 out of 126 (92 %) positive PCR samples and 40 out of 44 (91 %) dermatophyte-positive cultures. Implementation of PCR increased species-specific detection of dermatophytes by 21.1 %, leading to a threefold increase as compared to culture alone. Multiplex PCR offers a time-saving diagnostic tool for tinea unguium and augments laboratory assistance to clinical evaluation for proper treatment.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.079962-0
2015-01-01
2020-01-27
Loading full text...

Full text loading...

/deliver/fulltext/jmm/64/1/25.html?itemId=/content/journal/jmm/10.1099/jmm.0.079962-0&mimeType=html&fmt=ahah

References

  1. Alexander C. L., Shankland G. S., Carman W., Williams C.. ( 2011;). Introduction of a dermatophyte polymerase chain reaction assay to the diagnostic mycology service in Scotland. . Br J Dermatol 164:, 966–972. [CrossRef][PubMed]
    [Google Scholar]
  2. Beifuss B., Bezold G., Gottlöber P., Borelli C., Wagener J., Schaller M., Korting H. C.. ( 2011;). Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer. . Mycoses 54:, 137–145. [CrossRef][PubMed]
    [Google Scholar]
  3. Bergman A., Heimer D., Kondori N., Enroth H.. ( 2013;). Fast and specific dermatophyte detection by automated DNA extraction and real-time PCR. . Clin Microbiol Infect 19:, E205–E211. [CrossRef][PubMed]
    [Google Scholar]
  4. Bergmans A. M. C., van der Ent M., Klaassen A., Böhm N., Andriesse G. I., Wintermans R. G. F.. ( 2010;). Evaluation of a single-tube real-time PCR for detection and identification of 11 dermatophyte species in clinical material. . Clin Microbiol Infect 16:, 704–710. [CrossRef][PubMed]
    [Google Scholar]
  5. Bontems O., Hauser P. M., Monod M.. ( 2009;). Evaluation of a polymerase chain reaction-restriction fragment length polymorphism assay for dermatophyte and nondermatophyte identification in onychomycosis. . Br J Dermatol 161:, 791–796. [CrossRef][PubMed]
    [Google Scholar]
  6. Bossuyt P. M., Reitsma J. B., Bruns D. E., Gatsonis C. A., Glasziou P. P., Irwig L. M., Moher D., Rennie D., de Vet H. C. W., Lijmer J. G.. ( 2003;). The STARD statement for reporting studies of diagnostic accuracy: explanation and elaboration. . Clin Chem 49:, 7–18. [CrossRef][PubMed]
    [Google Scholar]
  7. Brasch J., Beck-Jendroschek V., Gläser R.. ( 2011;). Fast and sensitive detection of Trichophyton rubrum in superficial tinea and onychomycosis by use of a direct polymerase chain reaction assay. . Mycoses 54:, e313–e317. [CrossRef][PubMed]
    [Google Scholar]
  8. Brillowska-Dabrowska A., Saunte D. M., Arendrup M. C.. ( 2007;). Five-hour diagnosis of dermatophyte nail infections with specific detection of Trichophyton rubrum. . J Clin Microbiol 45:, 1200–1204. [CrossRef][PubMed]
    [Google Scholar]
  9. Brillowska-Dabrowska A., Nielsen S. S., Nielsen H. V., Arendrup M. C.. ( 2010;). Optimized 5-hour multiplex PCR test for the detection of tinea unguium: performance in a routine PCR laboratory. . Med Mycol 48:, 828–831. [CrossRef][PubMed]
    [Google Scholar]
  10. Cafarchia C., Iatta R., Latrofa M. S., Gräser Y., Otranto D.. ( 2013;). Molecular epidemiology, phylogeny and evolution of dermatophytes. . Infect Genet Evol 20:, 336–351. [CrossRef][PubMed]
    [Google Scholar]
  11. Chandran N. S., Pan J. Y., Pramono Z. A., Tan H. H., Seow C. S.. ( 2013;). Complementary role of a polymerase chain reaction test in the diagnosis of onychomycosis. . Australas J Dermatol 54:, 105–108. [CrossRef][PubMed]
    [Google Scholar]
  12. Clayton Y., Midgley G.. ( 1989;). Identification of agents of superficial mycoses. . In Medical Mycology: a Practical Approach. 1–25, Edited by Evans E. G. V., Richardson M. D... Oxford:: IRL Press;.
    [Google Scholar]
  13. Dhib I., Fathallah A., Charfeddine I. B., Meksi S. G., Said M. B., Slama F., Zemni R.. ( 2012;). Evaluation of chitine synthase (CHS1) polymerase chain reaction assay in diagnosis of dermatophyte onychomycosis. . J Mycol Med 22:, 249–255. [CrossRef][PubMed]
    [Google Scholar]
  14. Dhib I., Fathallah A., Yaacoub A., Hadj Slama F., Said M. B., Zemni R.. ( 2014;). Multiplex PCR assay for the detection of common dermatophyte nail infections. . Mycoses 57:, 19–26. [CrossRef][PubMed]
    [Google Scholar]
  15. Dubljanin E., Colovic Calovski I., Vujcic I., Dzamic A., Arendrup M. C., Petersen R. F., Jensen R. H.. ( 2014;). Clinical evaluation of a T. rubrum-specific polymerase chain reaction and pandermatophyte polymerase chain reaction in the diagnosis of suspected onychomycosis in 183 Serbian patients. . Br J Dermatol [Epub ahead of print]. [CrossRef][PubMed]
    [Google Scholar]
  16. Effendy I., Lecha M., Feuilhade de Chauvin M., Di Chiacchio N., Baran R.. ( 2005;). Epidemiology and clinical classification of onychomycosis. . J Eur Acad Dermatol Venereol 19: (Suppl 1), 8–12. [CrossRef][PubMed]
    [Google Scholar]
  17. Elavarashi E., Kindo A. J., Kalyani J.. ( 2013;). Optimization of PCR–RFLP directly from the skin and nails in cases of dermatophytosis, targeting the ITS and the 18S ribosomal DNA regions. . J Clin Diagn Res 7:, 646–651.[PubMed]
    [Google Scholar]
  18. Garg J., Tilak R., Singh S., Gulati A. K., Garg A., Prakash P., Nath G.. ( 2007;). Evaluation of pan-dermatophyte nested PCR in diagnosis of onychomycosis. . J Clin Microbiol 45:, 3443–3445. [CrossRef][PubMed]
    [Google Scholar]
  19. Ghannoum M. A., Mukherjee P. K., Warshaw E. M., Evans S., Korman N. J., Tavakkol A.. ( 2013;). Molecular analysis of dermatophytes suggests spread of infection among household members. . Cutis 91:, 237–245.[PubMed]
    [Google Scholar]
  20. Gräser Y., El Fari M., Vilgalys R., Kuijpers A. F. A., De Hoog G. S., Presber W., Tietz H.. ( 1999;). Phylogeny and taxonomy of the family Arthrodermataceae (dermatophytes) using sequence analysis of the ribosomal ITS region. . Med Mycol 37:, 105–114. [CrossRef][PubMed]
    [Google Scholar]
  21. Gräser Y., Czaika V., Ohst T.. ( 2012;). Diagnostic PCR of dermatophytes–an overview. . J Dtsch Dermatol Ges 10:, 721–725. [CrossRef][PubMed]
    [Google Scholar]
  22. Gupta A. K., Zaman M., Singh J.. ( 2008;). Diagnosis of Trichophyton rubrum from onychomycotic nail samples using polymerase chain reaction and calcofluor white microscopy. . J Am Podiatr Med Assoc 98:, 224–228. [CrossRef][PubMed]
    [Google Scholar]
  23. Hay R. J.. ( 1995;). Dermatophytosis and other superficial mycoses. . In Principles and Practice of Infectious Diseases, 261–283, , 4th edn.. Edited by Mandel G. L., Bennett J. E., Dolin R... New York:: Churchill Livingstone;.
    [Google Scholar]
  24. Idriss M. H., Khalil A., Elston D.. ( 2013;). The diagnostic value of fungal fluorescence in onychomycosis. . J Cutan Pathol 40:, 385–390. [CrossRef][PubMed]
    [Google Scholar]
  25. Jensen R. H., Arendrup M. C.. ( 2012;). Molecular diagnosis of dermatophyte infections. . Curr Opin Infect Dis 25:, 126–134. [CrossRef][PubMed]
    [Google Scholar]
  26. Kim J. Y., Choe Y. B., Ahn K. J., Lee Y. W.. ( 2011;). Identification of dermatophytes using multiplex polymerase chain reaction. . Ann Dermatol 23:, 304–312. [CrossRef][PubMed]
    [Google Scholar]
  27. Kondori N., Abrahamsson A. L., Ataollahy N., Wennerås C.. ( 2010;). Comparison of a new commercial test, Dermatophyte-PCR kit, with conventional methods for rapid detection and identification of Trichophyton rubrum in nail specimens. . Med Mycol 48:, 1005–1008. [CrossRef][PubMed]
    [Google Scholar]
  28. Li X. F., Tian W., Wang H., Chen H., Shen Y. N., Lv G. X., Liu W. D.. ( 2011;). Direct detection and differentiation of causative fungi of onychomycosis by multiplex polymerase chain reaction-based assay. . Eur J Dermatol 21:, 37–42.[PubMed]
    [Google Scholar]
  29. Litz C. E., Cavagnolo R. Z.. ( 2010;). Polymerase chain reaction in the diagnosis of onychomycosis: a large, single-institute study. . Br J Dermatol 163:, 511–514. [CrossRef][PubMed]
    [Google Scholar]
  30. Luk N. M., Hui M., Cheng T. S., Tang L. S., Ho K. M.. ( 2012;). Evaluation of PCR for the diagnosis of dermatophytes in nail specimens from patients with suspected onychomycosis. . Clin Exp Dermatol 37:, 230–234. [CrossRef][PubMed]
    [Google Scholar]
  31. Mehlig L., Garve C., Ritschel A., Zeiler A., Brabetz W., Weber C., Bauer A.. ( 2014;). Clinical evaluation of a novel commercial multiplex-based PCR diagnostic test for differential diagnosis of dermatomycoses. . Mycoses 57:, 27–34. [CrossRef][PubMed]
    [Google Scholar]
  32. Miyajima Y., Satoh K., Uchida T., Yamada T., Abe M., Watanabe S., Makimura M., Makimura K.. ( 2013;). Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes. . J Dermatol Sci 69:, 229–235. [CrossRef][PubMed]
    [Google Scholar]
  33. Moriarty B., Hay R., Morris-Jones R.. ( 2012;). The diagnosis and management of tinea. . BMJ 345:, e4380. [CrossRef][PubMed]
    [Google Scholar]
  34. Nenoff P., Herrmann J., Gräser Y.. ( 2007;). Trichophyton mentagrophytes sive interdigitale? A dermatophyte in the course of time. . J Dtsch Dermatol Ges 5:, 198–202. [CrossRef][PubMed]
    [Google Scholar]
  35. Pankewitz F., Nenoff P., Uhrlaß S., Bezold G., Winter I., Gräser Y.. ( 2013;). Development of a novel polymerase chain reaction-enzyme-linked immunosorbent assay for the diagnosis of Trichophyton rubrum onychomycosis. . Br J Dermatol 168:, 1236–1242. [CrossRef][PubMed]
    [Google Scholar]
  36. Paugam A., L’Ollivier C., Viguié C., Anaya L., Mary C., de Ponfilly G., Ranque S.. ( 2013;). Comparison of real-time PCR with conventional methods to detect dermatophytes in samples from patients with suspected dermatophytosis. . J Microbiol Methods 95:, 218–222. [CrossRef][PubMed]
    [Google Scholar]
  37. Rothmund G., Sattler E. C., Kaestle R., Fischer C., Haas C. J., Starz H., Welzel J.. ( 2013;). Confocal laser scanning microscopy as a new valuable tool in the diagnosis of onychomycosis - comparison of six diagnostic methods. . Mycoses 56:, 47–55. [CrossRef][PubMed]
    [Google Scholar]
  38. Salgo P. L., Daniel C. R., Gupta A. K., Mozena J. D., Joseph S. W.. ( 2003;). Onychomycosis disease management. . Medical Crossfire: J Debates, Peer Exchange and Insights in Medicine 4:, 1–17.
    [Google Scholar]
  39. Sato T., Takayanagi A., Nagao K., Tomatsu N., Fukui T., Kawaguchi M., Kudoh J., Amagai M., Yamamoto N., Shimizu N.. ( 2010;). Simple PCR-based DNA microarray system to identify human pathogenic fungi in skin. . J Clin Microbiol 48:, 2357–2364. [CrossRef][PubMed]
    [Google Scholar]
  40. Tsoumani M., Jelastopulu E., Bartzavali C., Vamvakopoulou S., Dimitracopoulos G., Anastassiou E. D., Christofidou M.. ( 2011;). Changes of dermatophytoses in southwestern Greece: an 18-year survey. . Mycopathologia 172:, 63–67. [CrossRef][PubMed]
    [Google Scholar]
  41. Verrier J., Pronina M., Peter C., Bontems O., Fratti M., Salamin K., Schürch S., Gindro K., Wolfender J. L.. & other authors ( 2012;). Identification of infectious agents in onychomycoses by PCR-terminal restriction fragment length polymorphism. . J Clin Microbiol 50:, 553–561. [CrossRef][PubMed]
    [Google Scholar]
  42. Verrier J., Krähenbühl L., Bontems O., Fratti M., Salamin K., Monod M.. ( 2013;). Dermatophyte identification in skin and hair samples using a simple and reliable nested polymerase chain reaction assay. . Br J Dermatol 168:, 295–301. [CrossRef][PubMed]
    [Google Scholar]
  43. Wisselink G. J., van Zanten E., Kooistra-Smid A. M.. ( 2011;). Trapped in keratin; a comparison of dermatophyte detection in nail, skin and hair samples directly from clinical samples using culture and real-time PCR. . J Microbiol Methods 85:, 62–66. [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.079962-0
Loading
/content/journal/jmm/10.1099/jmm.0.079962-0
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