Loop-Mediated Isothermal Amplification for the Diagnosis of Sporotrichosis by Sporothrix brasiliensis

IF 3.6 3区生物学 Q2 MYCOLOGY

Mycopathologia Pub Date : 2024-08-03 DOI:10.1007/s11046-024-00880-z

Marcella Donadel Araujo, Vanice Rodrigues Poester, Helena Schirmer, Vanessa Suñe Mattevi, Mariana Rodrigues Trápaga, Melissa Orzechowski Xavier, Marcus de Melo Teixeira, Rodrigo Almeida-Paes, Rosely Maria Zancopé-Oliveira, Cecília Bittencourt Severo

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Abstract

We aimed to develop and validate a Loop-mediated Isothermal Amplification (LAMP) assay to Sporothrix brasiliensis. LAMP reaction was developed using six primers designed based on calmodulin gene. In the LAMP reaction, we tested twenty isolates of S. brasiliensis from animals and humans, along with ten tissue samples extracted from the left footpad of mice that had been experimentally infected with S. brasiliensis. In addition, it included DNA samples from various other fungal species for specificity evaluation. All S. brasiliensis isolates yielded positive results in the LAMP, and the limit of DNA detection was 1 ng/μL. All murine samples were positive in the test while DNA from other fungal species were all negative, resulting in 100% of sensitivity and specificity of primers. LAMP diagnosis technique is a promising alternative to sporotrichosis diagnosis, in a simple and cost-effective way. Further studies are warranted to validate this technique using animal model samples obtained from both humans and animals.

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环路介导等温扩增用于诊断巴西孢子丝虫引起的孢子丝虫病

我们的目的是开发并验证一种针对巴西黑孢子虫的环介导等温扩增（LAMP）检测方法。我们使用基于钙调蛋白基因设计的六种引物开发了 LAMP 反应。在 LAMP 反应中，我们检测了从动物和人类身上分离出的二十个 S. brasiliensis，以及从实验感染 S. brasiliensis 的小鼠左足垫提取的十个组织样本。此外，它还包括其他各种真菌的 DNA 样本，用于特异性评估。在 LAMP 检测中，所有 S. brasiliensis 分离物均呈阳性，DNA 检测限为 1 ng/μL。所有鼠类样本的检测结果均为阳性，而其他真菌物种的DNA检测结果均为阴性，引物的灵敏度和特异性均为100%。LAMP 诊断技术是孢子丝菌病诊断的一种很有前途的替代方法，既简单又经济。有必要开展进一步研究，利用从人类和动物身上获得的动物模型样本来验证这一技术。

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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.

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