Johnson Matthew C, Damon Inger K, Karem Kevin L
Poxvirus Program, Division of Viral and Rickettsial Diseases, US Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, USA.
J Virol Methods. 2008 Jun;150(1-2):14-20. doi: 10.1016/j.jviromet.2008.02.009. Epub 2008 Apr 2.
Virus neutralization remains a vital tool in assessment of vaccine efficacy for smallpox in the absence of animal smallpox models. In this regard, development of a rapid, sensitive, and high-throughput vaccinia neutralization assay has been sought for evaluating alternative smallpox vaccines, use in bridging studies, as well as understanding the effects of anti-viral immunotherapeutic regimes. The most frequently used method of measuring vaccinia virus neutralization by plaque reduction is time, labor, and material intensive, and therefore limiting in its utility for large scale, high-throughput analysis. Recent advances provide alternative methods that are less labor intensive and higher throughput but with limitations in reagents needed and ease of use. An innovative neutralization assay is described based on a modified Western Reserve vaccinia vector expressing green fluorescent protein (WR-GFP) and an adherent cell monolayer in multi-well plate format. The assay is quick, accurate, provides a large dynamic range and is well suited for large-scale vaccination studies using standard adherent cell lines.
在缺乏动物天花模型的情况下，病毒中和仍然是评估天花疫苗效力的重要工具。在这方面，人们一直在寻求开发一种快速、灵敏且高通量的痘苗中和试验，用于评估替代天花疫苗、在桥接研究中的应用以及了解抗病毒免疫治疗方案的效果。最常用的通过蚀斑减少来测量痘苗病毒中和的方法耗时、费力且耗材，因此在大规模、高通量分析中的实用性有限。最近的进展提供了替代方法，这些方法劳动强度较低且通量较高，但在所需试剂和易用性方面存在局限性。本文描述了一种创新的中和试验，该试验基于表达绿色荧光蛋白的改良Western Reserve痘苗载体（WR-GFP）和多孔板形式的贴壁细胞单层。该试验快速、准确，提供了较大的动态范围，非常适合使用标准贴壁细胞系进行大规模疫苗接种研究。
