title: Performance of the Real Fungus-ID kit based on multiplex RT-PCR assay for the rapid detection and identification of Trichophyton spp. and Microsporum spp. in clinical specimens with suspected dermatophyte infection.
abstract: OBJECTIVE
  The aim of this study was to evaluate the performance of a commercially available multiplex RT-PCR assay for the rapid detection and identification of dermatophytes directly from clinical samples and cultures.
  
  
  RESULTS
  The multiplex RT-PCR assay was used to evaluate 118 clinical isolates from various specimen types and a total of 140 known specimens were compared with both conventional methods, commercially available PCR-REBA, and ITS sequence analysis. In this study, multiplex RT-PCR assay yield significantly more positive results than culture (91·9 vs 39·5%) and conventional methods including KOH microscopy (91·9 vs 71·3%). Although the results among the multiplex RT-PCR, PCR-REBA and ITS sequence analysis were concordant (100%) in 118 clinical isolates, concordant results between multiplex RT-PCR assay and culture were at 66% (78/118). The overall positive rates for the PCR-REBA, multiplex RT-PCR assay and ITS sequence analysis were 98·8, 91·9, and 52·9% respectively. In addition, the concordance rate of multiplex RT-PCR assay and the PCR-REBA assay was 93% (95% confidence interval (CI), 89·9-96·1, P < 0·0001), 93·7% (95% CI, 90·5-96·4, P < 0·0001) sensitivity, 100% (95% CI, 80·0-100, P < 0·0001) specificity, and 99·6% positive and 81% negative predictive values, respectively. Among the 258 samples, the most frequently identified dermatophyte species were Trichophyton rubrum (n = 199, 77·1%) and Trichophyton mentagrophytes (n = 28, 10·9%).
  
  
  CONCLUSIONS
  The entire multiplex RT-PCR procedure takes about 3 h, while results from culture can take up to 2-3 weeks. The use of the multiplex RT-PCR molecular diagnostic assay was rapid and reliable for detecting pathogen infections.
  
  
  CONCLUSIONS
  Even though the use of molecular diagnostic technology is more expensive than conventional methods, the clinical and economic benefit of saving time relative to expense remains to be elucidated. Therefore, the multiplex RT-PCR assay may provide the essential information to accelerate therapeutic decisions for earlier and adequate antibiotic treatment in the acute phase of fungal pathogen infections.
authors.affiliation: Wonju Eco Environmental Technology Center, M&D, Inc., Wonju, Korea.
authors.forename: H-Y
authors.initials: HY
authors.lastname: Wang
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: H
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Dermatology, Yonsei University Wonju College of Medicine, Wonju, Korea.
authors.forename: E H
authors.initials: EH
authors.lastname: Choi
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: H
authors.initials: H
authors.lastname: Lee
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D004271:DNA, Fungal / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008864:Microsporum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012871:Skin Diseases / Q000175:diagnosis / Q000382:microbiology*; D014249:Trichophyton / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D055815:Young Adult
chemical_list: D004271:DNA, Fungal
keywords: ITS sequence analysis; KOH microscopy; Microsporum spp.; PCR-REBA; Trichophyton spp; culture; diagnosis; multiplex RT-PCR