Kolokoltsov Andrey A, Wang Eryu, Colpitts Tonya M, Weaver Scott C, Davey Robert A
Department of Microbiology and Immunology, Center for Emerging Infectious Diseases, University of Texas Medical Branch, Galveston, TX 77555, USA.
Am J Trop Med Hyg. 2006 Oct;75(4):702-9.
Virus envelope proteins are the primary targets of neutralizing antibody responses. The epitopes recognized differ sufficiently between virus subtypes and species to distinguish viruses and provide an important basis for disease diagnosis. Venezuelan equine encephalitis virus (VEEV) causes acute febrile illness in humans and has high mortality in equines. The most specific detection methods for serum antibodies use live virus in neutralization assays or in blocking enzyme linked immunosorbent assays. However, work with Venezuelan equine encephalitis virus requires biosafety level 3 containment and select agent security in the United States. We report two new assays for detection of Venezuelan equine encephalitis virus neutralizing antibody responses, based on virus pseudotypes. The first provides detection by marker gene expression after 20 hours and is particularly suited for high-throughput screening; the second uses a new, rapid virus entry assay to give readouts within 1 hour. Both assays are safe, sensitive, and in general recapitulate neutralizing antibody titers obtained by conventional plaque reduction assays. Each is suitable as a rapid primary screen for detection of neutralizing antibodies against Venezuelan equine encephalitis virus.
病毒包膜蛋白是中和抗体反应的主要靶点。不同病毒亚型和物种之间识别的表位差异足够大，可用于区分病毒，并为疾病诊断提供重要依据。委内瑞拉马脑炎病毒（VEEV）可导致人类急性发热性疾病，在马类中具有高死亡率。血清抗体最特异的检测方法是在中和试验或阻断酶联免疫吸附试验中使用活病毒。然而，在美国，处理委内瑞拉马脑炎病毒需要生物安全3级防护和特定病原体安全措施。我们报告了两种基于病毒假型检测委内瑞拉马脑炎病毒中和抗体反应的新试验。第一种通过20小时后的标记基因表达进行检测，特别适合高通量筛选；第二种使用一种新的快速病毒进入试验，可在1小时内得出结果。这两种试验都安全、灵敏，并且总体上概括了通过传统蚀斑减少试验获得的中和抗体滴度。每种试验都适合作为检测针对委内瑞拉马脑炎病毒的中和抗体的快速初步筛选方法。
