Funding sources None.
CLINICAL AND LABORATORY INVESTIGATIONS
Development of a novel polymerase chain reaction–enzyme-linked immunosorbent assay for the diagnosis of Trichophyton rubrum onychomycosis
Article first published online: 5 JUN 2013
© 2013 The Authors. BJD © 2013 British Association of Dermatologists
British Journal of Dermatology
Volume 168, Issue 6, pages 1236–1242, June 2013
How to Cite
Pankewitz, F., Nenoff, P., Uhrlaß, S., Bezold, G., Winter, I. and Gräser, Y. (2013), Development of a novel polymerase chain reaction–enzyme-linked immunosorbent assay for the diagnosis of Trichophyton rubrum onychomycosis. British Journal of Dermatology, 168: 1236–1242. doi: 10.1111/bjd.12221
Conflicts of interest None declared.
- Issue published online: 5 JUN 2013
- Article first published online: 5 JUN 2013
- Accepted manuscript online: 10 JAN 2013 12:48AM EST
- Accepted for publication 27 December 2012
Background The prevalence of onychomycosis has increased steadily in the past decade. An accurate diagnosis at the outset is important for successful and cost-effective treatment of patients. However, current diagnostic tests for onychomycosis are not rapid, sensitive or specific.
Objectives To develop a microsatellite-based polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (MS-ELISA) for the detection of Trichophyton rubrum, which is the most common aetiological agent of onychomycosis.
Methods An archival set of 434 nail and skin specimens from 217 patients was included as the test sample in this study. We also compared MS-ELISA with an earlier published topoisomerase PCR-ELISA (TI-ELISA) using template DNA extracted by another method.
Results The MS-ELISA detected the highest number of positive samples (69%) followed by direct microscopy (56%), TI-ELISA (44%) and fungal culture (30%). When an identical DNA extraction method was applied to 120 specimens, the MS-ELISA proved to be twice as sensitive as the TI-ELISA.
Conclusions We have optimized a target gene and DNA extraction method for rapid detection of T. rubrum onychomycosis.