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应用基于微球的免疫测定法检测特定的大西洋鲑鱼抗鲑鱼甲病毒抗体。

Detection of specific Atlantic salmon antibodies against salmonid alphavirus using a bead-based immunoassay.

机构信息

Faculty of Veterinary Medicine, Norwegian University of Life Sciences (NMBU), Oslo, Norway.

SalMar Farming AS, Trondheim, Norway.

出版信息

Fish Shellfish Immunol. 2020 Nov;106:374-383. doi: 10.1016/j.fsi.2020.07.055. Epub 2020 Jul 30.

DOI:10.1016/j.fsi.2020.07.055
PMID:32738513
Abstract

Salmonid alphavirus (SAV) is the etiological cause of pancreas disease (PD) in Atlantic salmon (Salmo salar). Several vaccines against SAV are in use, but PD still cause significant mortality and concern in European aquaculture, raising the need for optimal tools to monitor SAV immunity. To monitor and control the distribution of PD in Norway, all salmonid farms are regularly screened for SAV by RT-qPCR. While the direct detection of SAV is helpful in the early stages of infection, serological methods could bring additional information on acquired SAV immunity in the later stages. Traditionally, SAV antibodies are monitored in neutralization assays, but they are time-consuming and cumbersome, thus alternative assays are warranted. Enzyme-linked immunosorbent assays (ELISAs) have not yet been successfully used for anti-SAV antibody detection in aquaculture. We aimed to develop a bead-based immunoassay for SAV-specific antibodies. By using detergent-treated SAV particles as antigens, we detected SAV-specific antibodies in plasma collected from both a SAV challenge trial and a field outbreak of PD. Increased levels of SAV-specific antibodies were seen after most fish had become negative for viral RNA. The bead-based assay is time saving compared to virus neutralization assays, and suitable for non-lethal testing due to low sample size requirements. We conclude that the bead-based immunoassay for SAV antibody detection is a promising diagnostic tool to complement SAV screening in aquaculture.

摘要

鲑鱼甲肝病毒(SAV)是大西洋鲑鱼(Salmo salar)胰腺疾病(PD)的病因。已经有几种针对 SAV 的疫苗在使用,但 PD 仍然在欧洲水产养殖业引起重大死亡率和关注,这就需要优化工具来监测 SAV 免疫。为了监测和控制挪威 PD 的分布,所有的鲑鱼养殖场都定期通过 RT-qPCR 筛查 SAV。虽然直接检测 SAV 有助于感染的早期阶段,但血清学方法可以在后期阶段提供关于获得性 SAV 免疫的额外信息。传统上,SAV 抗体通过中和测定来监测,但这些方法耗时且繁琐,因此需要替代测定。酶联免疫吸附测定(ELISA)尚未成功用于水产养殖中的抗 SAV 抗体检测。我们旨在开发一种用于检测 SAV 特异性抗体的基于珠子的免疫测定。通过使用经去污剂处理的 SAV 颗粒作为抗原,我们在来自 SAV 攻毒试验和 PD 现场暴发的血浆中检测到了 SAV 特异性抗体。在大多数鱼类的病毒 RNA 检测呈阴性后,SAV 特异性抗体的水平增加。与病毒中和测定相比,基于珠子的测定节省时间,并且由于需要的样本量较小,适合非致死性测试。我们得出结论,基于珠子的 SAV 抗体检测免疫测定是一种很有前途的诊断工具,可以补充水产养殖中的 SAV 筛选。

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