An automated assay platform for the evaluation of antiviral compounds against polioviruses

IF 2.2 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-08-08 DOI:10.1016/j.jviromet.2024.115006

Eric E. Rhoden, Bernardo A. Mainou

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Abstract

High-throughput screening requires assays that have flexibility to test large numbers of specimens while being accurate to ensure reproducibility across all specimens and variables tested. Previously, we used a low-throughput, cell-based assay to identify compounds with antiviral activity against polioviruses. In this report, we report the development and implementation of a high-throughput automation platform for the identification of compounds with antiviral activity against polioviruses. The platform uses off-the-shelf automated equipment combined with a modified assay, with minimal changes to existing laboratory space. We evaluated automation systems from Hudson Robotics Inc., Agilent Technologies, and a microplate reader from PerkinElmer during the platform design. Optimization for high throughput was focused on bulk reagent additions, serial dilutions, microplate washing and measuring results from the tens-to-hundreds of microplates. We evaluated the automated cell-based assay for selectivity, sensitivity, accuracy, precision, and reproducibility. This platform can be applied to screen novel antivirals against polioviruses and non-polio enteroviruses.

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评估抗脊髓灰质炎病毒化合物的自动测试平台。

高通量筛选要求检测方法既能灵活地检测大量样本，又能准确地确保所有样本和检测变量的可重复性。此前，我们曾使用一种基于细胞的低通量试验来鉴定对脊髓灰质炎病毒具有抗病毒活性的化合物。在本报告中，我们报告了用于鉴定对脊髓灰质炎病毒具有抗病毒活性的化合物的高通量自动化平台的开发和实施情况。该平台使用现成的自动化设备，结合改进的检测方法，对现有实验室空间的改动极小。在平台设计过程中，我们评估了哈德逊机器人公司（Hudson Robotics Inc）、安捷伦科技公司（Agilent Technologies）的自动化系统和珀金埃尔默公司（PerkinElmer）的微孔板阅读器。对高通量的优化主要集中在大量试剂添加、系列稀释、微孔板清洗以及测量数十到数百个微孔板的结果。我们对基于细胞的自动测试的选择性、灵敏度、准确性、精确性和可重复性进行了评估。该平台可用于筛选针对脊髓灰质炎病毒和非脊髓灰质炎肠道病毒的新型抗病毒药物。

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.