Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA), Rua Sarmento Leite, 285 - Centro Histórico, Porto Alegre, RS, 90050-170, Brasil.
Programa de Pós-Graduação Em Ciências da Saúde, Faculdade de Medicina (FAMED), Universidade Federal Do Rio Grande (FURG), Rio Grande, RS, Brasil.
Mycopathologia. 2024 Aug 3;189(5):73. doi: 10.1007/s11046-024-00880-z.
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.
我们旨在开发和验证一种针对巴西利蒙球孢子菌的环介导等温扩增（LAMP）检测方法。LAMP 反应是使用基于钙调蛋白基因设计的六个引物开发的。在 LAMP 反应中，我们测试了来自动物和人类的二十株巴西利蒙球孢子菌的分离株，以及从实验感染巴西利蒙球孢子菌的小鼠左脚垫提取的十个组织样本。此外，还包括用于特异性评估的各种其他真菌物种的 DNA 样本。所有巴西利蒙球孢子菌分离株在 LAMP 中均产生阳性结果，DNA 检测的下限为 1ng/μL。所有小鼠样本在检测中均为阳性，而来自其他真菌物种的 DNA 均为阴性，这导致引物的灵敏度和特异性均为 100%。LAMP 诊断技术是一种有前途的孢子丝菌病诊断替代方法，方法简单且具有成本效益。需要进一步的研究来使用从人和动物获得的动物模型样本验证该技术。
