Dual quantitative PCR assays for the rapid detection of Trichophyton indotineae from clinical samples

Dual quantitative PCR assays for the rapid detection of Trichophyton indotineae from clinical samples

Trichophyton indotineae is an emerging species of the Trichophyton mentagrophytes complex (TMC), responsible for an epidemic of widespread hairless skin infections that is frequently (50-70%) resistant to terbinafine. In order to initiate appropriate treatment as quickly as possible without waiting...

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Bibliographic Details
Published in:Medical mycology (Oxford) Vol. 62; no. 7
Main Authors: Baron, Audrey, Hamane, Samia, Gits-Muselli, Maud, Legendre, Lina, Benderdouche, Mazouz, Mingui, Anselme, Ghelfenstein-Ferreira, Théo, Alanio, Alexandre, Dellière, Sarah
Format: Journal Article
Language:English
Published: England 04-07-2024
Subjects:
Trichophyton indotineae
rapid molecular diagnosis
quantitative PCR
terbinafine resistance
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Summary:Trichophyton indotineae is an emerging species of the Trichophyton mentagrophytes complex (TMC), responsible for an epidemic of widespread hairless skin infections that is frequently (50-70%) resistant to terbinafine. In order to initiate appropriate treatment as quickly as possible without waiting for culture positivity (10-15 days) and molecular identification from the strain, we developed a dual quantitative PCR (qPCR) for the direct detection of T. indotineae in clinical samples. We first designed a T. indotineae specific qPCR assay (TI-qPCR) targeting a single specific polymorphism in the internal transcribed spacer region. Although none of the 94 non-dermatophyte and 7 dermatophyte species were amplified, this TI-qPCR allowed amplification of other TMC species at a lower yield. With equal amounts (0.1 ng) of DNA per reaction, the mean quantitative cycle (Cq) values for T. indotineae and non-indotineae TMC were 27.9 (±0.1) and 38.9 (±0.3), respectively. Therefore, we normalised this assay against a previously validated pan-dermatophyte qPCR assay (PD-qPCR) and relied on the ΔCq [(TI-qPCR) - (PD-qPCR)] to identify T. indotineae versus other TMC species. Dual assay was validated using 86 clinical samples of culture-confirmed T. indotinea and 19 non-indotineae TMC cases. The mean ΔCq for non-indotineae TMC was 9.6 ± 2.7, whereas the ΔCq for T. indotinea was -1.46 ± 2.1 (p < 0.001). Setting the ΔCq at 4.5 as a cut-off value resulted in 100% specificity for the detection of T. indotineae. This dual qPCR assay quickly detects T. indotineae from skin scrapings, aiding in early diagnosis and treatment for patients with suspected infection.
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ISSN:1369-3786
1460-2709
1460-2709
DOI:10.1093/mmy/myae067