Oceguera Leopoldo F, Patiris Peter J, Chiles Robert E, Busch Michael P, Tobler Leslie H, Hanson Carl V
California Department of Health Services, Viral and Rickettsial Disease Laboratory, Richmond, California 94804, USA.
Am J Trop Med Hyg. 2007 Jul;77(1):159-63.
The spread of West Nile virus (WNV) across the United States into areas with endemic flavivirus activity has complicated serologic surveillance of seasonal virus activity and diagnosis of infected individuals. Here we describe preliminary results from a comparison of serologic assays for flaviviruses: the reference plaque reduction neutralization test (PRNT), enzyme immunoassay (EIA), and a Western blot (WB) in which crude viral lysates were electrophoresed and blotted onto nitrocellulose. Human and chicken sera were tested and compared by each method against WNV and St. Louis encephalitis virus (SLEV). Antibody binding to three viral proteins determined WB interpretation: non-structural protein 1 (NS1), envelope (E), and pre-membrane (prM). WB results for a group of serially collected human plasma samples from WNV seroconverting blood donors were also correlated with transcription mediated amplification (TMA) and polymerase chain reaction (RT-PCR) results. Reactivity with NS1 appeared to be the most useful differentiating marker of WNV and SLEV infection in humans and chickens. Envelope protein was highly cross-reactive and, as indicated by additional results from dengue virus (DENV)-positive human sera, is perhaps useful serologically as a flavivirus group antigen.
西尼罗河病毒（WNV）在美国传播至有地方性黄病毒活动的地区，这使得季节性病毒活动的血清学监测以及感染个体的诊断变得复杂。在此，我们描述了黄病毒血清学检测方法比较的初步结果：参考蚀斑减少中和试验（PRNT）、酶免疫测定（EIA）以及一种蛋白质印迹法（WB），其中粗制病毒裂解物经电泳后转移至硝酸纤维素膜上。采用每种方法对人和鸡的血清进行检测，并针对WNV和圣路易斯脑炎病毒（SLEV）进行比较。抗体与三种病毒蛋白的结合情况决定了WB的解读结果：非结构蛋白1（NS1）、包膜蛋白（E）和前膜蛋白（prM）。一组来自WNV血清阳转献血者的连续采集的人血浆样本的WB结果也与转录介导扩增（TMA）和聚合酶链反应（RT-PCR）结果相关。与NS1的反应性似乎是人和鸡中WNV和SLEV感染最有用的鉴别标志物。包膜蛋白具有高度交叉反应性，并且正如登革病毒（DENV）阳性人血清的其他结果所示，它在血清学上可能作为黄病毒属抗原是有用的。
