evaluation of multiplex real time pcr for identifying dermatophytes in clinical samples- a multicenter study

• Main Author: Binsky, Inbal I
• Format: Web Resource
• Language: English
• Published: Morressier 01.01.2017
• DOI: 10.26226/morressier.5ac3832d2afeeb00097a4929

Background: Most superficial fungal infections are caused by dermatophytes. Conventional methods of dermatophyte detection have inherent disadvantages. Advanced molecular-technology-based methods hold promise for the rapid and accurate diagnosis of dermatophytoses directly from clinical samples. Materials/methods: 4 multicenter evaluations were performed; A total of 604 clinical samples (261+200+103+40) with suspected fungal infection were obtained from the Dermatology labs.Extracted DNA from skin scale speciment, plucked hair, nails, culture and swabs of human origin was detected on a real time PCR- based multiplex application for fast, reliable, ready to use and high throughput in vitro diagnostic of dermatophytes. This multiplex PCR system enables 7 different dermatophytes types detection within 2 hours. The test contains two ready to use mixes for detection of 7 most common dermatopytes types: [ 1 ] Trichophyton tonsurans [ 2 ] Trichophyton violaceum [ 3 ] Trichophyton rubrum complex [ 4 ] Trichophyton mentagrophytes complex [ 5 ] Microsporum canis [ 6 ] Microsporum gypseum [ 7 ] Epidermophyton floccosum and Internal Control.The dermatophyte detection rate was compared to KOH smear and culture. Inconsistencies between the methods were resolved by sequencing. Results: The RT-PCR results including DNA extractions were available within 4 hours. There are between 10-30% false negative results of KOH and culture compared to PCR, meaning PCR method has highly sensetivity and specificity compared to common methods.tCUL+KOH+tCUL+KOH-tCUL-KOH+tCUL-KOH-PCR+t254t24t51t52PCR-t3t4t19t197Conclusions: The DERMADYN kit based on multiplex RT-PCR platform is a rapid, more sensitive and efficient method for identifying dermatophyte species in clinical samples. Our findings justify its use as a first step in the diagnostic algorithm of superficial fungal infections.