Varea R, Monleón E, Pacheco C, Luján L, Bolea R, Vargas M A, Van Eynde G, Saman E, Dickson L, Harkiss G, Amorena B, Badiola J J
Department of Animal Pathology, University of Zaragoza, Veterinary Faculty, Spain.
J Vet Diagn Invest. 2001 Jul;13(4):301-7. doi: 10.1177/104063870101300404.
The aim of this work was to investigate whether an enzyme-linked immunosorbent assay (ELISA) was useful for early detection of maedi-visna virus (MVV) infection in sheep under field conditions. An ELISA based on p25 recombinant protein and a gp46 synthetic peptide was used. Sequentially obtained serum samples (n = 1,941) were studied for 4 years. ELISA results were compared with those of the agar gel immunodiffusion (AGID) test, and results of both tests were compared with a reference result established using consensus scores for at least 2 of 3 serologic techniques (AGID, ELISA, and western blotting, which was used to resolve result discrepancies between the other 2 techniques). A total of 247 discrepancies were observed between ELISA and AGID. Of these, 131 were due to an earlier detection of 120 sera by the ELISA and 11 sera by AGID. The remaining discrepancies (116) were due to the presence of false reactions in both tests. Fewer false-negative results were found by ELISA than with AGID (6 vs. 69 sera, respectively), whereas the number of false-positive results was virtually the same for ELISA and AGID (21 vs. 20, respectively). In relation to the reference result, ELISA sensitivity and specificity were 97.8% and 98.2%, respectively, whereas values for AGID were 76.3% and 98.3%, respectively. The agreement between ELISA and the reference result was higher than that between AGID and the reference result (K value: 0.96 and 0.77, respectively). A variation in the ELISA signal (based on optical density) was observed during the study period, suggesting different antibody levels throughout the animal's life. The ELISA was useful for detecting MVV-infected sheep in field conditions and has potential for use in control and eradication programs.
本研究的目的是调查酶联免疫吸附测定(ELISA)是否有助于在野外条件下早期检测绵羊梅迪-维斯纳病毒(MVV)感染。使用了基于p25重组蛋白和gp46合成肽的ELISA。连续4年对1941份血清样本进行了研究。将ELISA结果与琼脂凝胶免疫扩散(AGID)试验结果进行比较,并将两种试验的结果与使用三种血清学技术(AGID、ELISA和western blotting,用于解决其他两种技术之间的结果差异)中至少两种的共识评分建立的参考结果进行比较。ELISA和AGID之间共观察到247处差异。其中,131处差异是由于ELISA比AGID更早检测到120份血清和11份血清。其余差异(116处)是由于两种试验中均存在假反应。ELISA发现的假阴性结果比AGID少(分别为6份血清和69份血清),而ELISA和AGID的假阳性结果数量基本相同(分别为21份和20份)。相对于参考结果,ELISA的敏感性和特异性分别为97.8%和98.2%,而AGID的值分别为76.3%和98.3%。ELISA与参考结果之间的一致性高于AGID与参考结果之间的一致性(K值分别为0.96和0.77)。在研究期间观察到ELISA信号(基于光密度)存在变化,表明动物一生中抗体水平不同。ELISA可用于在野外条件下检测感染MVV的绵羊,具有用于控制和根除计划的潜力。