Antibody Response to Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Nonstructural Proteins and Implications for Diagnostic Detection and Differentiation of PRRSV Types I and II
Antibody Response to Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Nonstructural Proteins and Implications for Diagnostic Detection and Differentiation of PRRSV Types I and II
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DOI：
10.1128/cvi.00483-08
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发表时间：
2009-05-01
影响因子：
--
通讯作者：
Fang, Ying
中科院分区：
文献类型：
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作者：
Brown, Elizabeth;Lawson, Steven;Fang, Ying
为了进一步描述猪对猪繁殖与呼吸综合征病毒（PRRSV）的体液免疫应答，采用直接酶联免疫吸附试验（ELISA）研究了实验性暴露于病毒的猪体内针对PRRSV非结构蛋白的抗体应答动力学。最高的免疫反应性是对nsp 1，nsp 2和nsp 7。利用重组nsp 7作为抗原，我们验证了双重ELISA用于同时检测和区分针对I型和II型PRRSV的血清抗体。基于1,334例已知阳性和1,357例已知阴性样本的受试者工作特征分析显示出良好的特异性（I型98.3%，II型99.3%）和敏感性（I型97.4%，II型99.8%）。为了区分I型和II型PRRSV，对470份来自实验接种猪的血清进行了测试，在470份样品中的469份中正确区分了阳性血清。测定了nsp 7双重ELISA检测来自感染各种遗传上不同的田间菌株的猪的血清抗体应答的能力。nsp 7双重ELISA与Idexx HerdChek PRRS 2XR ELISA具有97.6%的一致性。在Idexx ELISA疑似假阳性样本的进一步检测中，nsp 7双重ELISA将98%的样本解析为阴性。综上所述，这些结果表明nsp 7双重ELISA可用作PRRSV血清学的鉴别测试，具有高水平的灵敏度和特异性。该ELISA为常规或后续诊断提供了额外的工具，并在流行病学调查和疫情调查中具有重要价值。
To further characterize the humoral immune response of pigs to porcine reproductive and respiratory syndrome virus ( PRRSV), direct enzyme-linked immunosorbent assays (ELISA) were used to study the kinetics of antibody responses directed against PRRSV nonstructural proteins in pigs experimentally exposed to the virus. The highest immunoreactivities were against nsp1, nsp2, and nsp7. Using the recombinant nsp7 as an antigen, we validated a dual ELISA for the simultaneous detection and differentiation of serum antibodies against type I and type II PRRSV. Receiver operating characteristic analysis based on 1,334 known-positive and 1,357 known-negative samples showed good specificity (98.3% to type I and 99.3% to type II) and sensitivity (97.4% for type I and 99.8% for type II). To differentiate type I and type II PRRSV, 470 sera originating from experimentally inoculated pigs were tested, and positive sera were correctly differentiated in 469 of 470 samples. The capability of the nsp7 dual ELISA to detect serum antibody responses from pigs infected with various genetically different field strains was determined. The nsp7 dual ELISA possessed 97.6% agreement with the Idexx HerdChek PRRS 2XR ELISA. In further testing of Idexx ELISA suspected false-positive samples, the nsp7 dual ELISA resolved 98% of the samples as negative. Taken together, these results indicate that the nsp7 dual ELISA can be used as a differential test for PRRSV serology with high levels of sensitivity and specificity. This ELISA offers an additional tool for routine or follow-up diagnostics, as well as having substantial value in epidemiological surveys and outbreak investigations.