Development of a Vertical Flow Dot-Immunobinding Assay (Dot-Iba) for Rapid Detection of Neoscytalidium dimidiatum.

IF 4.1 3区生物学 Q2 MYCOLOGY

Mycopathologia Pub Date : 2026-03-10 DOI:10.1007/s11046-026-01066-5

Akkarapong Plengpanich, Sumanas Bunyaratavej, Anchalee Tungtrongchitr, Nawannaporn Saelim, Thapani Srisai, Charussri Leeyaphan, Piriyaporn Chongtrakool, Pichet Ruenchit

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Abstract

Neoscytalidium dimidiatum is a non-dermatophyte mold that commonly causes skin and nail infections in tropical regions and often resists conventional antifungal therapies. Because its clinical and laboratory features often resemble dermatophyte infections, diagnosis is frequently delayed and treatment is sometimes inappropriate. We therefore developed a dot-immunobinding assay (Dot-Iba) to detect N. dimidiatum antigens. We generated a highly specific monoclonal antibody, 3E6F7 (MAb 3E6F7), for antigen capture, and used goat anti-mouse Ig conjugated with alkaline phosphatase (AP) as the signal generator. The test pad comprised a test hole, a nitrocellulose membrane (NC), and water-absorbent pads in a vertical flow-through format to allow a rapid antigen-antibody reaction. The assembled system detected N. dimidiatum antigens in vitro with high specificity and yielded visible results within 2 h; its detection limit was 0.9 µg without cross-reactivity to dermatophyte or non-dermatophyte fungi. This rapid, specific, and easy-to-use assay shows strong potential as a diagnostic tool, particularly in settings with limited access to fungal culture or advanced molecular diagnostics, where early, accurate identification is crucial.

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垂直流动点免疫结合法（Dot-Iba）快速检测新西塔利菌的建立。

新锡塔利菌是一种非皮肤真菌霉菌，通常引起皮肤和指甲感染在热带地区，往往抵抗传统的抗真菌治疗。由于其临床和实验室特征往往类似于皮肤真菌感染，诊断经常被延误，治疗有时是不适当的。因此，我们开发了一种斑点免疫结合试验（Dot-Iba）来检测双斑棘球蚴抗原。我们制备了一种高特异性单克隆抗体3E6F7 （MAb 3E6F7）用于抗原捕获，并使用山羊抗小鼠Ig偶联碱性磷酸酶（AP）作为信号发生器。测试垫包括一个测试孔、一个硝化纤维素膜（NC）和垂直流动形式的吸水垫，以允许快速的抗原-抗体反应。组装后的系统体外检测双斑棘球蚴抗原，特异性高，2 h内检测结果明显；检出限为0.9µg，对皮生菌和非皮生菌均无交叉反应。这种快速、特异且易于使用的检测显示出作为诊断工具的巨大潜力，特别是在真菌培养或高级分子诊断有限的环境中，早期、准确的鉴定至关重要。

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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.