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一种用于评估抗脊髓灰质炎病毒化合物的自动化分析平台。

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

机构信息

Polio and Picornavirus Branch, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.

Polio and Picornavirus Branch, Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.

出版信息

J Virol Methods. 2024 Sep;329:115006. doi: 10.1016/j.jviromet.2024.115006. Epub 2024 Aug 8.

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.

摘要

高通量筛选需要具有灵活性的测定法,能够测试大量样本,同时确保所有样本和测试变量的可重复性。以前,我们使用一种低通量的基于细胞的测定法来鉴定具有抗脊髓灰质炎病毒活性的化合物。在本报告中,我们报告了开发和实施一种用于鉴定具有抗脊髓灰质炎病毒活性的化合物的高通量自动化平台。该平台使用现成的自动化设备结合改良的测定法,对现有实验室空间的改动最小。在平台设计过程中,我们评估了 Hudson Robotics Inc.、Agilent Technologies 和 PerkinElmer 的自动化系统。针对高通量进行了优化,重点是批量试剂添加、连续稀释、微孔板清洗以及从数十到数百个微孔板测量结果。我们评估了自动化基于细胞的测定法的选择性、灵敏度、准确性、精密度和重现性。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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/47d3/11881792/2d1b0c769342/nihms-2057907-f0001.jpg

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