Lu Taofeng, Wang Yuanzhi, Wu Yanjun, Zhao Lili, Wu Shuguang, Chen Hongyan
Institute for Laboratory Animal Research, Guizhou University of Traditional Chinese Medicine, Guiyang, 550025, China.
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 678 Haping Road, Harbin, 150069, China.
Arch Virol. 2021 Jan;166(1):83-90. doi: 10.1007/s00705-020-04850-w. Epub 2020 Oct 17.
Aleutian mink disease (AMD), caused by Aleutian mink disease virus (AMDV), is a very important infectious disease of mink. Currently, elimination of antibody- or antigen-positive animals is the most successful strategy for eradicating AMD, but the claw-cutting method of blood sampling is difficult to perform and painful for the animal. In this study, we aimed to establish an antigen capture enzyme-linked immunosorbent assay (AC-ELISA) method for the efficient detection of AMDV antigens using fecal samples. A purified mouse monoclonal antibody (mAb) was used as the capture antibody, and a rabbit polyclonal antibody (pAb) was used as the detection antibody. The assay was optimized by adjusting a series of parameters. Using a cutoff value of 0.205, the limit of detection of the AC-ELISA for strain AMDV-G antigen was 2 μg/mL, and there was no cross-reaction with other mink viruses. The intra- and inter-assay standard deviations were below 0.046, and the correlation of variance (CV) values were 1.24-7.12% when testing fecal samples. Compared with conventional PCR results, the specificity and sensitivity were 91.5% and 90.6%, respectively, and the concordance rate between the two methods was 91.1%.
阿留申水貂病（AMD）由阿留申水貂病病毒（AMDV）引起，是水貂的一种非常重要的传染病。目前，淘汰抗体或抗原阳性动物是根除AMD最成功的策略，但剪爪采血方法操作困难且会给动物带来痛苦。在本研究中，我们旨在建立一种抗原捕获酶联免疫吸附测定（AC-ELISA）方法，用于使用粪便样本高效检测AMDV抗原。使用纯化的小鼠单克隆抗体（mAb）作为捕获抗体，兔多克隆抗体（pAb）作为检测抗体。通过调整一系列参数对该测定进行了优化。使用0.205的临界值，AC-ELISA对AMDV-G株抗原的检测限为2μg/mL，与其他水貂病毒无交叉反应。检测粪便样本时，批内和批间标准差均低于0.046，变异系数（CV）值为1.24 - 7.12%。与传统PCR结果相比，特异性和敏感性分别为91.5%和90.6%，两种方法的符合率为91.1%。
