In vitro Assay to Assess Efficacy of Potential Antiviral Compounds against Enterovirus D68
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Author:
Date:
2017-03-20
[Abstract] In 2014 enterovirus D68 (EV-D68) caused the largest outbreak in the United States since the discovery of the virus. Distinct from before, the 2014 infections were associated with more severe respiratory disease and occasional neurological complications. So far, there are no available vaccines or antivirals for the prophylaxis or treatment of EV-D68 infections. In order to evaluate the antiviral activity of potential inhibitors of EV-D68 replication, a cell-based cytopathic effect (CPE) reduction assay was developed (Sun et al., 2015).
[摘要] 2014年肠病毒D68(EV-D68)自发现病毒以来,在美国引起了最大的爆发。与之前不同的是,2014年感染与更严重的呼吸道疾病和偶尔的神经系统并发症有关。到目前为止,还没有可用的疫苗或抗病毒药物用于预防或治疗EV-D68感染。为了评估EV-D68复制潜在抑制剂的抗病毒活性,开发了基于细胞的细胞病变效应(CPE)降低测定法(Sun等,2015)。
背景 作为新出现的病原体,以前很少报道靶向EV-D68的抗病毒化合物。迫切需要建立和开发抗病毒方法来对抗潜在的EV-D68流行病。在这里,我们报告一个详细的方案,可用于识别选择性抗EV-D68化合物。为了筛选和鉴定潜在的抗病毒化合物,基于MTS的CPE降解测定法是可重复使用的,易于使用和省时的方法,被广泛使用。该方法依赖于化合物抑制EV-D68诱导的CPE降低的能力。当化合物主动抑制EV-D68的复制时,病毒诱导的CPE将被减少或不存在。因此,代谢活性细胞能够将黄色四唑盐底物(MTS / PMS)转化为棕色甲an制品。当化合物不具有抗病毒作用时,宿主细胞死于病毒诱导的CPE,其将缺乏代谢活性,并且黄色底物保持未代谢。比色转化是定量的,因为可以从该测定产生的数据计算EC 50。
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Infectious Virus Yield Assay for Hepatitis E Virus
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Author:
Date:
2014-08-05
[Abstract] Hepatitis E virus (HEV) is one of the main causes of acute hepatitis worldwide. Infections are particularly severe in pregnant women and chronic hepatitis E is known to occur in immunocompromised patients. Current therapy (ribavirin or pegylated alpha interferon) has severe side effects and cannot be employed in all patients. In order to evaluate potential new inhibitors of HEV replication, a virus yield assay can be employed in which the amount of viral RNA progeny released into the culture medium is quantified by reverse-transcription quantitative PCR (RT-qPCR) (Debing et al., 2014).
[摘要] 戊型肝炎病毒(HEV)是全世界急性肝炎的主要原因之一。 感染在孕妇中特别严重,已知慢性戊型肝炎发生在免疫受损的患者中。 目前的治疗(利巴韦林或聚乙二醇化α干扰素)具有严重的副作用,不能用于所有患者。 为了评估潜在的新的HEV复制抑制剂,可以使用病毒产量测定法,其中释放到培养基中的病毒RNA子代的量通过逆转录定量PCR(RT-qPCR)(Debing等人, al 。,2014)。
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Luminescence-based Antiviral Assay for Hepatitis E Virus
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Author:
Date:
2014-08-05
[Abstract] Hepatitis E virus (HEV) is one of the main causes of acute hepatitis worldwide. Infections are particularly severe in pregnant women and chronic hepatitis E is known to occur in immunocompromised patients. Current therapy (ribavirin or pegylated alpha interferon) has severe side effects and cannot be employed in all patients. In order to evaluate potential new inhibitors of HEV replication, a luminescence-based replicon assay is particularly useful since it offers a rapid read-out and does not pose any biosafety risks (Debing et al., 2014).
[摘要] 戊型肝炎病毒(HEV)是全世界急性肝炎的主要原因之一。 感染在孕妇中特别严重,已知慢性戊型肝炎发生在免疫受损的患者中。 目前的治疗(利巴韦林或聚乙二醇化α干扰素)具有严重的副作用,不能用于所有患者。 为了评价潜在的新的HEV复制抑制剂,基于发光的复制子测定法是特别有用的,因为其提供了快速读出,并且不会引起任何生物安全风险(Debing等人,2014) 。
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