Development of an antigen-capture enzyme-linked immunosorbent assay for diagnosis of Aleutian mink disease virus
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ORIGINAL ARTICLE
Development of an antigen‑capture enzyme‑linked immunosorbent assay for diagnosis of Aleutian mink disease virus Taofeng Lu1 · Yuanzhi Wang2 · Yanjun Wu1 · Lili Zhao2 · Shuguang Wu1 · Hongyan Chen2 Received: 26 April 2020 / Accepted: 6 September 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract 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%.
Introduction Aleutian mink disease (AMD), caused by Aleutian mink disease virus (AMDV) [1], is a very important contagious disease in the mink-breeding industry worldwide. AMDV has been assigned by the International Committee on Taxonomy of Viruses (ICTV) [2, 3] to the genus Amdoparvovirus, subfamily Parvovirinae, family Parvoviridae. Two structural proteins (VP1 and VP2) and three nonstructural proteins (NS1, NS2, and NS3) [4, 5] are encoded by the AMDV genome. The VP2 protein is the major immunogenic Handling Editor: Ana Cristina Bratanich. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00705-020-04850-w) contains supplementary material, which is available to authorized users. * Hongyan Chen [email protected] 1
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
2
protein and is involved in viral tropism, host selection, and pathogenicity [6–8]. AMDV is a major pathogen that restricts the development of the mink-breeding industry in China, and there is no effective vaccine against the disease [9, 10]. The seropositive rate was as high as 68.67% in major mink-breeding areas of China (Heilongjiang, Liaoning, Jilin, Hebei and Shandong provinces) during the period of 2009–2011 [11, 12]. The host range of
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