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Evaluation of two commercial multiplex real-time PCR assays for the detection of dermatophytes when onychomycosis or tinea pedis is suspected
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
2016 (English)Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
Abstract [en]

Onychymycosis and tinea pedis are fungal infections in humans caused by dermatophytes. The high prevalence of onychomycosis and the difficulties associated with its treatment increases the costs for healthcare significantly and new high quality methods for the detection of dermatophytes is needed. Presently, two methods are utilized for detection of these infections, microscopy and cultivation. Microscopy is a method with low specificity and sensitivity that demands an experienced user and the possibility to detect different species is lacking. Cultivation can differentiate between species but is time consuming since it takes 1 up to 4 weeks to grow the fungi on agar plates. Recently, molecular methods has been developed for the detection of dermatophytes. In this study, two commercial multiplex real-time PCR assays for determination of dermatophytes were tested for evaluation in the clinical routine laboratory, DermaGenius Nail Multiplex kit and RIDA®GENE Dermatophytes kit and compared to the in-house conventional PCR method. The methods used in this study includes DNA-extraction of clinical samples followed by extraction in two steps, thermal lysis DNA-extraction followed by a DNA-extraction with Bullet. Thereafter the samples were analyzed using multiplex realtime-PCR. The results indicate that the DermaGenius® kit had higher sensitivity and specificity compared to the in-house PCR kit and is therefore suitable to use for routine analysis at the clinical microbiology laboratory. The RIDA®GENE Dermatophyte kit had a much lower sensitivity than the other two methods, possibly explained by the fact that it is still under development by the manufacturer.

Place, publisher, year, edition, pages
2016.
Keywords [en]
Trichophyton rubrum, molecular biology, DermaGenius, RIDA®GENE, SSI Diagnostica
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:uu:diva-349022OAI: oai:DiVA.org:uu-349022DiVA, id: diva2:1199244
Educational program
Biomedical Laboratory Science Programme; Biomedical Laboratory Science Programme
Examiners
Available from: 2018-04-20 Created: 2018-04-19 Last updated: 2018-04-20Bibliographically approved

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CiteExportLink to record
Permanent link

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Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
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Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
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Output format
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