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Detection of Serum Antibodies to Ovine Progressive Pneumonia Virus in Sheep by Using a Caprine Arthritis-Encephalitis Virus Competitive-Inhibition Enzyme-Linked Immunosorbent Assay

机译:山羊关节炎-脑炎病毒竞争抑制酶联免疫吸附法检测绵羊绵羊进展性肺炎病毒血清抗体

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摘要

A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) for detection of antibodies to the surface envelope (SU) of caprine arthritis-encephalitis virus (CAEV) was recently reported (L. M. Herrmann, W. P. Cheevers, T. C. McGuire, D. Scott Adams, M. M. Hutton, W. G. Gavin, and D. P. Knowles, Clin. Diagn. Lab. Immunol. 10:267-271, 2003). The cELISA utilizes CAEV-63 SU captured on microtiter plates using the monoclonal antibody (MAb) F7-299 and measures competitive displacement of binding of the anti-CAEV MAb GPB 74A by goat serum. The present study evaluated the CAEV cELISA for detection of antibodies to ovine progressive pneumonia virus (OPPV) in sheep. Three hundred thirty-two sera were randomly selected from 21,373 sheep sera collected throughout the United States to determine the sensitivity and specificity of cELISA and agar gel immunodiffusion (AGID) based on immunoprecipitation (IP) of [35S]methionine-labeled OPPV antigens as a standard of comparison. A positive cELISA test was defined as >20.9 percent inhibition (% I) of MAb 74A binding based on two standard deviations above the mean % I of 191 IP-negative sheep sera. At this cutoff, there were 2 of 141 false-negative sera (98.6% sensitivity) and 6 of 191 false-positive sera (96.9% specificity). Sensitivity and specificity values for IP-monitored AGID were comparable to those for cELISA for 314 of 332 sera with unambiguous AGID results. Concordant results by cELISA and IP resolved 16 of the 18 sera that were indeterminate by AGID. Additional studies evaluated cELISA by using 539 sera from a single OPPV-positive flock. Based on IP of 36 of these sera, there was one false-negative by cELISA among 21 IP-positive sera (95.5% sensitivity) and 0 of 15 false-positives (100% specificity). We conclude that the CAEV cELISA can be applied to detection of OPPV antibodies in sheep with high sensitivity and specificity.
机译:最近报道了一种竞争抑制酶联免疫吸附试验(cELISA),用于检测针对山羊关节炎-脑炎病毒(CAEV)的表面包膜(SU)的抗体(LM Herrmann,WP Cheevers,TC McGuire,D。Scott Adams, MM Hutton,WG Gavin,和DP Knowles,Clin.Diagn.Lab.Immunol.10:267-271,2003)。 cELISA利用使用单克隆抗体(MAb)F7-299在微量滴定板上捕获的CAEV-63 SU来测量山羊血清中抗CAEV MAb GPB 74A结合的竞争性置换。本研究评估了CAEV cELISA用于检测绵羊绵羊进行性肺炎病毒(OPPV)抗体的能力。从美国全境收集的21,373只绵羊血清中随机选择三百二十二份血清,以基于[35S]蛋氨酸标记的OPPV抗原的免疫沉淀(IP)的方法测定cELISA和琼脂凝胶免疫扩散(AGID)的敏感性和特异性。比较标准。基于高于191个IP阴性绵羊血清的平均I%的两个标准差,将阳性cELISA测试定义为对单克隆抗体74A结合的抑制> 20.9%(I)。在此截止值时,在141个假阴性血清中有2个(灵敏度为98.6%),在191个假阳性血清中有6个(特异性为96.9%)。 IP监测的AGID的灵敏度和特异性值可与332血清中314血清的cELISA的灵敏度和特异性值相媲美,而AGID结果无歧义。 cELISA和IP的一致结果可解析18份血清中的16份,由AGID不确定。其他研究通过使用来自单个OPPV阳性羊群的539血清评估了cELISA。基于这些血清中36种的IP,通过cELISA检测到21种IP阳性血清中有1种假阴性(敏感性为95.5%),而15种假阳性中有0种为阴性(100%特异性)。我们得出的结论是,CAEV cELISA可用于检测绵羊OPPV抗体,具有很高的灵敏度和特异性。

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