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

H. Y. Wang, H. Kim, Eung Ho Choi, Hyeyoung Lee

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Abstract

Aims: 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. Methods and 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. Significance and Impact of the Study: 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.

Original languageEnglish
Pages (from-to)234-247
Number of pages14
JournalJournal of Applied Microbiology
Volume120
Issue number1
DOIs
Publication statusPublished - 2016 Jan 1

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Microsporum
Arthrodermataceae
Trichophyton
Multiplex Polymerase Chain Reaction
Fungi
Infection
Sequence Analysis
Polymerase Chain Reaction
Molecular Pathology
Confidence Intervals
Mycoses
Microscopy
Economics
Anti-Bacterial Agents
Technology

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

@article{c2d8d259bdf24e2ea928af0597cfe679,
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 = "Aims: 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. Methods and 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. Significance and Impact of the Study: 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.",
author = "Wang, {H. Y.} and H. Kim and Choi, {Eung Ho} and Hyeyoung Lee",
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T1 - 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

AU - Wang, H. Y.

AU - Kim, H.

AU - Choi, Eung Ho

AU - Lee, Hyeyoung

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Aims: 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. Methods and 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. Significance and Impact of the Study: 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.

AB - Aims: 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. Methods and 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. Significance and Impact of the Study: 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.

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