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肉汤微量稀释体外筛选:一种检测新型抗真菌化合物的简便快速方法。

Broth Microdilution In Vitro Screening: An Easy and Fast Method to Detect New Antifungal Compounds.

作者信息

de-Souza-Silva Calliandra Maria, Guilhelmelli Fernanda, Zamith-Miranda Daniel, de Oliveira Marco Antônio, Nosanchuk Joshua Daniel, Silva-Pereira Ildinete, Albuquerque Patrícia

机构信息

Laboratory of Molecular Biology, Department of Cellular Biology, Institute of Biological Sciences, University of Brasília.

Department of Microbiology and Immunology, Albert Einstein College of Medicine; Division of Infectious Diseases, Department of Medicine, Albert Einstein College of Medicine.

出版信息

J Vis Exp. 2018 Feb 14(132):57127. doi: 10.3791/57127.

Abstract

Fungal infections have become an important medical condition in the last decades, but the number of available antifungal drugs is limited. In this scenario, the search for new antifungal drugs is necessary. The protocol reported here details a method to screen peptides for their antifungal properties. It is based on the broth microdilution susceptibility test from the Clinical and Laboratory Standards Institute (CLSI) M27-A3 guidelines with modifications to suit the research of antimicrobial peptides as potential new antifungals. This protocol describes a functional assay to evaluate the activity of antifungal compounds and may be easily modified to suit any particular class of molecules under investigation. Since the assays are performed in 96-well plates using small volumes, a large-scale screening can be completed in a short amount of time, especially if carried out in an automation setting. This procedure illustrates how a standardized and adjustable clinical protocol can help the bench-work pursuit of new molecules to improve the therapy of fungal diseases.

摘要

在过去几十年中,真菌感染已成为一种重要的医学病症,但可用的抗真菌药物数量有限。在这种情况下,寻找新的抗真菌药物是必要的。本文报道的方案详细介绍了一种筛选具有抗真菌特性肽的方法。它基于临床和实验室标准协会(CLSI)M27 - A3指南中的肉汤微量稀释药敏试验,并进行了修改以适应作为潜在新型抗真菌剂的抗菌肽研究。该方案描述了一种评估抗真菌化合物活性的功能测定方法,并且可以很容易地进行修改以适应所研究的任何特定类别的分子。由于测定是在96孔板中使用少量体积进行的,因此可以在短时间内完成大规模筛选,特别是在自动化环境中进行时。该程序说明了标准化且可调整的临床方案如何有助于在实验室工作中寻找新分子以改善真菌疾病的治疗。

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