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QuickFit:一种高通量基于 RT-qPCR 的检测方法,用于定量体外病毒生长和适应性。

QuickFit: A High-Throughput RT-qPCR-Based Assay to Quantify Viral Growth and Fitness In Vitro.

机构信息

Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, Harvard University, Cambridge, MA 02139, USA.

出版信息

Viruses. 2024 Aug 19;16(8):1320. doi: 10.3390/v16081320.

Abstract

Quantifying viral growth rates is key to understanding evolutionary dynamics and the potential for mutants to escape antiviral drugs. Defining evolutionary escape paths and their impact on viral fitness allows for the development of drugs that are resistant to escape. In the case of HIV, combination antiretroviral therapy can successfully prevent or treat infection, but it relies on strict adherence to prevent escape. Here, we present a method termed QuickFit that enables the quantification of viral fitness by employing large numbers of parallel viral cultures to measure growth rates accurately. QuickFit consistently recapitulated HIV growth measurements obtained by traditional approaches, but with significantly higher throughput and lower rates of error. This method represents a promising tool for rapid and consistent evaluation of viral fitness.

摘要

量化病毒增长率对于理解进化动态和突变体逃避抗病毒药物的潜力至关重要。定义进化逃逸途径及其对病毒适应性的影响,可以开发出能够抵抗逃逸的药物。在 HIV 的情况下,联合抗逆转录病毒疗法可以成功预防或治疗感染,但它依赖于严格的依从性来防止逃逸。在这里,我们提出了一种称为 QuickFit 的方法,该方法通过使用大量平行的病毒培养物来准确测量增长率,从而实现了对病毒适应性的量化。QuickFit 一致地再现了传统方法获得的 HIV 生长测量值,但具有更高的通量和更低的错误率。这种方法代表了一种快速和一致评估病毒适应性的有前途的工具。

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