Development of a novel polymerase chain reaction-enzyme-linked immunosorbent assay for the diagnosis of Trichophyton rubrum onychomycosis.

Pankewitz, F.; Nenoff, P.; Uhrlaß, S.; Bezold, G.; Winter, I.; Gräser, Y.

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.

POLYMERASE chain reaction; ENZYME-linked immunosorbent assay; DNA polymerases; TRICHOPHYTON; ONYCHOMYCOSIS; DISEASE prevalence; DIAGNOSIS

British Journal of Dermatology, 2013, Vol 168, Issue 6, p1236

0007-0963

Academic Journal

10.1111/bjd.12221