Burke D S, Tingpalapong M, Elwell M R, Paul P S, Van Deusen R A
Am J Vet Res. 1985 Oct;46(10):2054-7.
A solid-phase enzyme-linked immunosorbent assay (ELISA) was developed for detection of porcine immunoglobulin (Ig)M antibodies to Japanese encephalitis virus (JEV). Antibodies in sera were captured onto the solid phase of Microtiter plates sensitized with mouse monoclonal antibodies to porcine mu heavy chain. Virus antigen binding to the lawn of IgM was quantitated by subsequent binding of peroxidase-labeled human hyperimmune anti-JEV IgG, which in the final step, catalyzed a substrate color change. In sucrose density-gradient fractionated sera from recently infected pigs, the peak of ELISA JEV IgM activity corresponded to the peak of 18-S, 2-mercaptoethanol-sensitive hemagglutination-inhibiting (HAI) antibody activity. Within 2 to 3 days, JEV-infected sentinel pigs developed high JEV IgM activity; this activity decreased within 2 weeks. Among specimens collected from 99 random swine at abattoirs in Thailand during a period of low JEV transmission, none of 25 JEV HAI-negative sera had JEV IgM activity, 7 of 74 JEV HAI-positive sera did have JEV IgM activity, and the remaining 67 sera had readily detectable JEV HAI antibodies, but lacked JEV IgM. The JEV IgM solid-phase ELISA was useful for rapidly diagnosing active or recent JEV infections in swine.
建立了一种固相酶联免疫吸附测定法（ELISA），用于检测猪抗日本脑炎病毒（JEV）的免疫球蛋白（Ig）M抗体。血清中的抗体被捕获到用抗猪μ重链小鼠单克隆抗体致敏的微量滴定板固相上。通过随后结合过氧化物酶标记的人超免疫抗JEV IgG来定量与IgM菌苔结合的病毒抗原，在最后一步中，该酶催化底物颜色变化。在最近感染猪的蔗糖密度梯度分级血清中，ELISA JEV IgM活性峰值与18-S、2-巯基乙醇敏感血凝抑制（HAI）抗体活性峰值相对应。在2至3天内，感染JEV的哨兵猪产生高JEV IgM活性；该活性在2周内下降。在JEV传播率较低期间，从泰国屠宰场的99头随机猪采集的样本中，25份JEV HAI阴性血清均无JEV IgM活性，74份JEV HAI阳性血清中有7份具有JEV IgM活性，其余67份血清具有易于检测到的JEV HAI抗体，但缺乏JEV IgM。JEV IgM固相ELISA可用于快速诊断猪的活动性或近期JEV感染。
