title: Development of a novel polymerase chain reaction-enzyme-linked immunosorbent assay for the diagnosis of Trichophyton rubrum onychomycosis.
abstract: 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.
  
  
  OBJECTIVE
  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.
authors.affiliation: Konsiliarlabor für Dermatophyten, Institut für Mikrobiologie und Hygiene, Universitätsmedizin-Berlin Charité, Dorotheenstr. 96, D-10117 Berlin, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Pankewitz
authors.forename: P
authors.initials: P
authors.lastname: Nenoff
authors.forename: S
authors.initials: S
authors.lastname: Uhrlaß
authors.forename: G
authors.initials: G
authors.lastname: Bezold
authors.forename: I
authors.initials: I
authors.lastname: Winter
authors.forename: Y
authors.initials: Y
authors.lastname: Gräser
mesh_terms: D000908:Antibodies, Fungal / Q000097:blood; D004271:DNA, Fungal / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005189:False Positive Reactions; D006801:Humans; D014009:Onychomycosis / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D014249:Trichophyton / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000908:Antibodies, Fungal; D004271:DNA, Fungal