Detection of Pan-Dermatophytes and Trichophyton rubrum Using Recombinase Polymerase Amplification-Lateral Flow Dipstick Assay.

IF 3.6 3区生物学 Q2 MYCOLOGY

Mycopathologia Pub Date : 2024-12-30 DOI:10.1007/s11046-024-00921-7

Qian Wang, Xin Huang, Qiuhong Yan, Ruikang Chen, Lifang Shao, Ruoyu Li, Yinggai Song, Xiaoying Yuan

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引用次数: 0

Abstract

Background: Traditional methods for diagnosing onychomycosis are characterized by limited sensitivity and prolonged processing times, and heavily rely on the skill level of laboratory personnel.

Objectives: To develop a fast, simple, user-friendly, and reliable molecular assay that offers high sensitivity and specificity for the detection of common dermatophytes in nail specimens.

Methods: We developed a technique that integrates recombinase polymerase isothermal amplification with lateral flow dipstick (RPA-LFD) for the detection of pan-dermatophytes and Trichophyton rubrum, and evaluated its analytical sensitivity and specificity. This method was applied to analyze 190 nail specimens, with the results compared with traditional microscopy and fungal culture.

Results: The RPA-LFD assay demonstrated an analytical sensitivity of 10 pg/reaction for pan-dermatophytes and 1 pg/reaction for T. rubrum. In clinical evaluations for tinea unguium, the sensitivity of the RPA-LFD, fungal culture, and microscopy methods, as determined through latent class analysis, was estimated to be 91.0%, 70.8%, and 93.9%, respectively. Correspondingly, the specificity of these methods-RPA-LFD, fungal culture, and microscopy-was assessed at approximately 95.7%, 98.0%, and 94.3%.

Conclusions: Our RPA-LFD assay exhibited high sensitivity and specificity in the detection of dermatophytes. Due to its technical simplicity, enhanced sensitivity, and reduced processing times, it represents a promising alternative to conventional fungal culture methods for the mycological detection and identification of dermatophytes.

查看原文本刊更多论文

重组酶聚合酶扩增-横向流动试纸法检测泛皮肤菌和红毛癣菌。

背景：传统的诊断甲真菌病的方法灵敏度有限，处理时间长，严重依赖实验室人员的技能水平。目的：建立一种快速、简单、用户友好、可靠的分子检测方法，为指甲标本中常见皮肤真菌的检测提供高灵敏度和特异性。方法：建立了重组酶聚合酶等温扩增与侧流试纸（RPA-LFD）相结合的泛皮肤菌和红毛癣菌检测技术，并对其分析灵敏度和特异性进行了评价。应用该方法对190例甲标本进行了分析，并与传统镜检和真菌培养结果进行了比较。结果：RPA-LFD法对泛皮菌的分析灵敏度为10 pg/反应，对红毛t的分析灵敏度为1 pg/反应。在临床评估中，RPA-LFD、真菌培养和显微镜方法的敏感性，通过潜在分类分析确定，估计分别为91.0%、70.8%和93.9%。相应地，这些方法（rpa - lfd、真菌培养和显微镜）的特异性分别为95.7%、98.0%和94.3%。结论：RPA-LFD法检测皮肤真菌具有较高的敏感性和特异性。由于其技术简单，灵敏度提高，处理时间缩短，它代表了传统真菌培养方法的一种有希望的替代方法，用于皮肤真菌的真菌学检测和鉴定。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Mycopathologia 生物-真菌学

CiteScore

6.80

自引率

3.60%

发文量

审稿时长

3 months

期刊介绍： Mycopathologia is an official journal of the International Union of Microbiological Societies (IUMS). Mycopathologia was founded in 1938 with the mission to ‘diffuse the understanding of fungal diseases in man and animals among mycologists’. Many of the milestones discoveries in the field of medical mycology have been communicated through the pages of this journal. Mycopathologia covers a diverse, interdisciplinary range of topics that is unique in breadth and depth. The journal publishes peer-reviewed, original articles highlighting important developments concerning medically important fungi and fungal diseases. The journal highlights important developments in fungal systematics and taxonomy, laboratory diagnosis of fungal infections, antifungal drugs, clinical presentation and treatment, and epidemiology of fungal diseases globally. Timely opinion articles, mini-reviews, and other communications are usually invited at the discretion of the editorial board. Unique case reports highlighting unprecedented progress in the diagnosis and treatment of fungal infections, are published in every issue of the journal. MycopathologiaIMAGE is another regular feature for a brief clinical report of potential interest to a mixed audience of physicians and laboratory scientists. MycopathologiaGENOME is designed for the rapid publication of new genomes of human and animal pathogenic fungi using a checklist-based, standardized format.