Serodiagnosis, targeting nonstructural protein 4, of porcine reproductive and respiratory syndrome virus
Serodiagnosis, targeting nonstructural protein 4, of porcine reproductive and respiratory syndrome virus
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针对猪繁殖与呼吸综合征病毒非结构蛋白 4 的血清诊断
DOI：
10.1007/s00705-017-3625-5
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发表时间：
2018-02-01
影响因子：
2.7
通讯作者：
Xiao, Yihong
中科院分区：
文献类型：
--
作者：
Cao, Shengliang;Cai, Xinna;Xiao, Yihong
猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus，PRRSV）是一种严重危害养猪业的病毒，给养猪业造成了巨大的经济损失。通常，N蛋白包被的间接酶联免疫吸附测定（N包被的iELISA）用于检测PRRSV抗体。非结构蛋白（NSP）4是PRRSV生命周期所必需的，含有B细胞表位。然而，在临床样品中没有检测到针对NSP 4的特异性抗体。在这项研究中，我们开发了NSP 4包被的iELISA，并比较其有效性与N包被的iELISA。通过测试一组70份PRRSV阳性和80份PRRSV阴性猪血清样品，开发了NSP 4包被的iELISA，在450 nm的光密度下具有0.406的截止值，其产生100%的特异性和灵敏度。NSP 4包被和N包被的iELISA之间的一致性为92.2%。有趣的是，50份血清样品，主要来自用HP-PRRSV活毒株接种的猪，用NSP 4包被的iELISA检测PRRSV抗体呈阳性，但用N包被的iELISA检测为阴性。这些结果通过蛋白质印迹分析和另一种基于NSP 2的N-末端的iELISA（NSP 2 -1-包被的iELISA）进一步证实。蛋白质印迹分析结果与NSP 4包被和NSP 2 -1包被的iELISA分析结果之间的一致性分别为92%和96.1%，表明所开发的NSP 4包被的iELISA是区分对HP-PRRSV疫苗的假阴性应答与真阴性应答的有用工具。
Porcine reproductive and respiratory syndrome virus (PRRSV) causes great economic losses to the swine industry worldwide. Typically, an N protein-coated indirect enzyme-linked immunosorbent assay (N-coated iELISA) is used to detect PRRSV antibodies. Non-structural protein (NSP) 4 is essential to the PRRSV life cycle and contains B-cell epitopes. Yet, no specific antibody against NSP4 has been detected in clinical samples. In this study, we developed an NSP4-coated iELISA and compared its effectiveness with the N-coated iELISA. The NSP4-coated iELISA was developed with a cut-off value of 0.406 at an optical density of 450 nm by testing a panel of 70 PRRSV positive and 80 PRRSV negative pig serum samples, which generated a specificity and sensitivity of 100%. Agreement between the NSP4-coated and N-coated iELISAs was 92.2%. Interestingly, 50 serum samples, mostly from pigs vaccinated with the HP-PRRSV live strain, tested positive for PRRSV antibodies with the NSP4-coated iELISA, but were negative with the N-coated iELISA. These results were further confirmed by western blot analysis and another iELISA based on the N-terminus of NSP2 (NSP2-1-coated iELISA). The agreement between the results of western blot analysis with the NSP4-coated and NSP2-1-coated iELISA analyses were 92% and 96.1%, respectively, showing that the developed NSP4-coated iELISA is a useful tool to discriminate a false negative from a true negative response to the HP-PRRSV vaccine.