Libeau G, Préhaud C, Lancelot R, Colas F, Guerre L, Bishop D H, Diallo A
CIRAD-EMVT, Laboratoire de Pathotrop, Maisons-Alfort, France.
Res Vet Sci. 1995 Jan;58(1):50-5. doi: 10.1016/0034-5288(95)90088-8.
A competitive ELISA based on the reaction between a monoclonal antibody (mAb) and a recombinant nucleoprotein of the peste des petits ruminants virus (PPRV) was developed. This protein was obtained in large quantities from insect cells infected with a PPR nucleoprotein recombinant baculovirus (N-B). The competitive ELISA was compared with the virus neutralisation test (VNT) for detecting specific antibodies to PPRV in sheep and goats. The time consuming VNT is the only prescribed test that is capable of distinguishing between PPRV and the cross-reactive rinderpest virus (RPV). The competitive ELISA involves the simultaneous addition of the mAb and antibodies present in a positive serum, leading to competition for a specific epitope on the N-B. Optimum conditions were obtained by using serum samples which had positive or negative neutralising activity against PPRV or RPV. A negative cut-off point was determined on PPRV-negative sera from RPV-vaccinated cattle. A threshold value of 48 per cent inhibition, calculated from the mean for this population plus 2.7 standard deviations, was used in routine testing. A total of 683 sera were analysed by the competitive ELISA and the VNT. A good correlation (r = 0.94) was observed between the titres obtained in the two tests, with 80 sera that were from laboratory sources. The agreement between the two tests was determined on 271 field sera (kappa = 0.825). Their relative sensitivity (94.5 per cent) and specificity (99.4 per cent) were assessed on the 148 laboratory sera plus the 271 sera used for the determination of kappa.(ABSTRACT TRUNCATED AT 250 WORDS)
基于单克隆抗体（mAb）与小反刍兽疫病毒（PPRV）重组核蛋白之间反应的竞争性酶联免疫吸附测定（ELISA）被开发出来。这种蛋白是从感染了PPR核蛋白重组杆状病毒（N - B）的昆虫细胞中大量获得的。将该竞争性ELISA与病毒中和试验（VNT）进行比较，以检测绵羊和山羊体内针对PPRV的特异性抗体。耗时的VNT是唯一规定的能够区分PPRV和交叉反应性牛瘟病毒（RPV）的试验。竞争性ELISA涉及同时加入mAb和阳性血清中存在的抗体，导致对N - B上特定表位的竞争。通过使用对PPRV或RPV具有阳性或阴性中和活性的血清样本获得了最佳条件。在接种了RPV疫苗的牛的PPRV阴性血清中确定了阴性临界值。在常规检测中使用从该群体平均值加上2.7个标准差计算得出的48%抑制阈值。总共683份血清通过竞争性ELISA和VNT进行了分析。在80份来自实验室的血清中，两种试验获得的滴度之间观察到良好的相关性（r = 0.94）。在271份现场血清上确定了两种试验之间的一致性（kappa = 0.825）。在148份实验室血清加上用于确定kappa的271份血清上评估了它们的相对敏感性（94.5%）和特异性（99.4%）。（摘要截断于250字）
