用酶联免疫吸附法检测猪水疱病病毒抗体的非传染性病毒样颗粒抗原

Young-Joon Ko, Kang-Seuk Choi, Jin-Ju Nah, David J Paton, Jae-Ku Oem, Ginette Wilsden, Shien-Young Kang, Nam-In Jo, Joo-Ho Lee, Jae-Hong Kim, Hee-Woo Lee, Jong-Myeong Park
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引用次数: 24

摘要

灭活的猪水疱病病毒(SVDV)抗原被用于目前的酶联免疫吸附试验(elisa),用于检测猪水疱病病毒(SVDV)的抗体。为了开发一种非感染性的重组载体,我们利用双杆状病毒重组体,在不同启动子下同时表达SVDV的P1和3CD蛋白基因,在形态和抗原性上与原SVDV相似,制备了SVDV样颗粒(VLPs)。5b7 ELISA试剂盒检测结果显示,重组VLPs与SVDV颗粒的抗原性差异无统计学意义,说明重组VLPs可以代替SVDV抗原在ELISA试剂盒中使用。我们利用VLPs和SVDV特异性中和单克隆抗体3H10 (VLP-ELISA)建立了阻断ELISA检测猪SVDV血清抗体。在猪SVDV中和阴性血清(n= 1041)中检测,VLP-ELISA具有99.9%的高特异性。当使用从实验感染三种不同SVDV毒株的猪(n=19)定期收集的血清(n=186)进行测试时,VLP-ELISA早在感染后3天就检测到SVDV血清抗体,并继续检测所有感染猪的抗体,直到实验结束(感染后121天)。该检测结果与金标准病毒中和试验结果相似,表明VLP-ELISA检测猪SVDV血清抗体具有较高的特异性和敏感性。本文首次报道了SVDV重组VLPs的制备及诊断应用。讨论了VLPs的进一步潜在用途。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Noninfectious virus-like particle antigen for detection of swine vesicular disease virus antibodies in pigs by enzyme-linked immunosorbent assay.

An inactivated SVDV antigen is used in current enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies to swine vesicular disease virus (SVDV). To develop a noninfectious recombinant alternative, we produced SVDV-like particles (VLPs) morphologically and antigenically resembling authentic SVDV particles by using a dual baculovirus recombinant, which expresses simultaneously the P1 and 3CD protein genes of SVDV under different promoters. Antigenic differences between recombinant VLPs and SVDV particles were not statistically significant in results obtained with a 5B7-ELISA kit, indicating that the VLPs could be used in the place of SVDV antigen in ELISA kits. We developed a blocking ELISA using the VLPs and SVDV-specific neutralizing monoclonal antibody 3H10 (VLP-ELISA) for detection of SVDV serum antibodies in pigs. The VLP-ELISA showed a high specificity of 99.9% when tested with pig sera that are negative for SVDV neutralization (n=1,041). When tested using sera (n=186) collected periodically from pigs (n=19) with experimental infection with each of three different strains of SVDV, the VLP-ELISA detected SVDV serum antibodies as early as 3 days postinfection and continued to detect the antibodies from all infected pigs until termination of the experiments (up to 121 days postinfection). This test performance was similar to that of the gold standard virus neutralization test and indicates that the VLP-ELISA is a highly specific and sensitive method for the detection of SVDV serum antibodies in pigs. This is the first report of the production and diagnostic application of recombinant VLPs of SVDV. Further potential uses of the VLPs are discussed.

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