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- Title
一种基于VP0 蛋白检测 DHAV(1 型和3 型)血清抗体的 间接 ELISA 的建立及应用.
- Authors
马波; 戚海惠; 张文晶; 陈浩田; 张琪; 常蕊; 刘悦; 张雪莲; 张桂红; 王君伟
- Abstract
In order to establish a rapid serological method for detecting the antibodies against Duck hepatitis A virus (DHAV) type 1 and type 3 (DHAV-1, DHAV-3), the reactionogenicities of recombinant proteins VP0, VP1 and VP3 with DHAV-1 and DHAV-3 were compared by using ELISA assays. The result showed that the DHAV-1 VP0 could react with serum antibodies against both DHAV-1 and DHAV-3 serving as a universal antigen. After the optimization of reaction conditions, an indirect ELISA was established based on DHAV-1 VP0 recombinant protein for the antibodies detection of DHAV-1, DHAV-3. The mainly optimized reaction conditions were as following: the concentration of coated antigen was 0.25 μg/mL, and the dilutions of serum and goat against duck HRP-IgG were 1:100 and 1:400 respectively. The specificity assay showed that the ELISA only detected antibodies against DHAV-1 and DHAV-3 but had no cross-reaction with other serum antibodies against goose parvovirus (GPV), duck enteritis virus (DEV), H9N2 subtype avian influenza virus (H9N2 AIV) and duck tambusu virus (DTMUV). The sensitivity was 1:320 of DHAV-3 positive serum, and the coefficient variability of intra-batch was less than 4% and inter-batch was less than 6%. Compared with commercial DHAV ELISA antibody detect kit, the coincidences of positive and negative rate were 93.3% and 88.9% , respectively, and the total coincidence rate was 95.7%. The results suggested that the indirect ELISA could detect the clinical DHAV serum samples. This study provid a technological method to detect and/or monitor DHAV serum antibody level for drafting effective preventative measure.
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2020, Vol 42, Issue 3, p245
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.201905031