一种用于评估抗爱泼斯坦-巴尔病毒抗病毒化合物的快速DNA杂交检测法。
A rapid DNA hybridization assay for the evaluation of antiviral compounds against Epstein-Barr virus.
作者信息
Prichard Mark N, Daily Shannon L, Jefferson Geraldine M, Perry Amie L, Kern Earl R
机构信息
Department of Pediatrics, University of Alabama School of Medicine, Birmingham, AL 35233, USA.
出版信息
J Virol Methods. 2007 Sep;144(1-2):86-90. doi: 10.1016/j.jviromet.2007.04.001. Epub 2007 May 30.
Abstract
There is a need for additional therapies for Epstein-Barr virus (EBV) infections, but the routine screening of large numbers of potential inhibitors has been difficult due to the laborious nature of traditional assays. A new rapid assay was developed to evaluate compounds for antiviral activity against this virus that is both rapid and robust. Test compounds are added to cultures of Akata cells in 96-well plates that have been induced to undergo a lytic infection. Viral DNA produced during the infection is transferred to a membrane and quantified using a non-radioactive DNA hybridization assay. This assay was validated using a set of compounds with known activity against EBV and results compared favorably to an established real-time PCR assay. Subsequent experience with this assay has confirmed that it offers improved efficiency and robustness compared to other assays used routinely to evaluate candidate compounds for antiviral activity against EBV.
摘要
对于爱泼斯坦-巴尔病毒（EBV）感染，需要有更多的治疗方法，但由于传统检测方法费力，对大量潜在抑制剂进行常规筛选一直很困难。人们开发了一种新的快速检测方法，用于评估化合物对这种病毒的抗病毒活性，该方法既快速又可靠。将测试化合物添加到已诱导发生裂解感染的96孔板中的赤羽病（Akata）细胞培养物中。感染过程中产生的病毒DNA转移到膜上，并使用非放射性DNA杂交检测法进行定量。使用一组对EBV具有已知活性的化合物对该检测方法进行了验证，结果与既定的实时PCR检测法相比具有优势。随后使用该检测方法的经验证实，与常规用于评估针对EBV抗病毒活性的候选化合物的其他检测方法相比，它具有更高的效率和可靠性。
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