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Generation characterization and application in serodiagnosis of recombinant swine vesicular disease virus-like particles

机译:重组猪水泡病病毒样颗粒的产生表征及在血清学诊断中的应用

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摘要

Swine vesicular disease (SVD) is a highly contagious viral disease that causes vesicular disease in pigs. The importance of the disease is due to its indistinguishable clinical signs from those of foot-and-mouth disease, which prevents international trade of swine and related products. SVD-specific antibody detection via an enzyme-linked immunosorbent assay (ELISA) is the most versatile and commonly used method for SVD surveillance and export certification. Inactivated SVD virus is the commonly used antigen in SVD-related ELISA. A recombinant SVD virus-like particle (VLP) was generated by using a Bac-to-Bac baculovirus expression system. Results of SVD-VLP analyses from electron microscopy, western blotting, immunofluorescent assay, and mass spectrometry showed that the recombinant SVD-VLP morphologically resemble authentic SVD viruses. The SVD-VLP was evaluated as a replacement for inactivated whole SVD virus in competitive and isotype-specific ELISAs for the detection of antibodies against SVD virus. The recombinant SVD-VLP assay produced results similar to those from inactivated whole virus antigen ELISA. The VLP-based ELISA results were comparable to those from the virus neutralization test for antibody detection in pigs experimentally inoculated with SVD virus. Use of the recombinant SVD-VLP is a safe and valuable alternative to using SVD virus antigen in diagnostic assays.
机译:猪水泡病(SVD)是一种高度传染性的病毒性疾病,会引起猪的水泡病。该疾病的重要性是由于其与口蹄疫没有明显的临床体征,从而阻止了猪及其相关产品的国际贸易。通过酶联免疫吸附测定(ELISA)检测SVD特异性抗体是用于SVD监视和出口认证的最通用,最常用的方法。灭活的SVD病毒是SVD相关ELISA中常用的抗原。通过使用Bac-to-Bac杆状病毒表达系统生成重组SVD病毒样颗粒(VLP)。通过电子显微镜,蛋白质印迹,免疫荧光分析和质谱分析得出的SVD-VLP分析结果表明,重组SVD-VLP在形态上类似于真实的SVD病毒。在竞争性和同种型特异性ELISA中,评估SVD-VLP作为灭活的完整SVD病毒的替代品,以检测针对SVD病毒的抗体。重组SVD-VLP分析产生的结果类似于灭活的全病毒抗原ELISA的结果。基于VLP的ELISA结果与通过病毒中和试验进行的SVD病毒实验猪抗体检测的结果相当。重组SVD-VLP的使用是在诊断分析中使用SVD病毒抗原的一种安全且有价值的替代方法。

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