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Establishment and Application of Indirect ELISAs for Detecting Antibodies against Goose Astrovirus Genotype 1 and 2

文献类型： 外文期刊

作者： Zhang, Mengran 1 ; Wei, Xinyu 1 ; Qian, Jing 1 ; Yu, Zhengyu 6 ; Liu, Xin 6 ; Luo, Yan 6 ; Zhang, Haitao 6 ; Gu, Youfang 2 ; Li, Yin 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Med, Nanjing 210014, Peoples R China

2.Anhui Sci & Technol Univ, Coll Anim Sci, Fengyang 233100, Peoples R China

3.Minist Agr, Key Lab Vet Biol Engn & Technol, Nanjing 210014, Peoples R China

4.GuoTai Taizhou Ctr Technol Innovat Vet Biol, Taizhou 225300, Peoples R China

5.Nanjing Agr Univ, Coll Vet Med, Key Lab Anim Dis Diagnost & Immunol, Minist Agr, Nanjing 210095, Peoples R China

6.Lihua Nanjing Agr Ind Res Inst Co Ltd, Nanjing 210014, Peoples R China

7.Jiangsu Lihua Anim Husb Co Ltd, Changzhou 213168, Peoples R China

关键词： goose astrovirus; GAstV-1; GAstV-2; capsid protein; antibody; indirect ELISA

期刊名称：VACCINES （ 影响因子：7.8； 五年影响因子：7.4 ）

ISSN：

年卷期： 2023 年 11 卷 3 期

页码：

收录情况： SCI

摘要： Goose astrovirus (GAstV) was classified into GAstV-1 and GAstV-2, and both caused gosling viral gout. Recently, there has been no effective commercial vaccine to control the infection. It is important to establish serological methods to distinguish between the two genotypes. In this study, we reported the development and application of two indirect enzyme-linked immunosorbent assays (ELISAs) using the GAstV-1 virus and a recombinant GAstV-2 capsid protein as specific antigens to detect antibodies against GAstV-1 and GAstV-2, respectively. The optimal coating antigen concentration of indirect GAstV-1-ELISA and GAstV-2-Cap-ELISA was 1.2 mu g/well and 125 ng/well, respectively. In addition, the antigen coating temperature and time, sera dilution and reaction time, and the dilution and reaction time of HRP-conjugated secondary antibody were optimized. The cut-off values were 0.315 and 0.305, and the analytical sensitivity was 1:6400 and 1:3200 for indirect GAstV-1-ELISA and GAstV-2-Cap-ELISA, respectively. The assays were able to differentiate specific sera against GAstVs, TUMV, GPV, and H9N2-AIV. The intra- and inter-plate variabilities of indirect ELISAs were less than 10%. The coincidence rate of positive sera was higher than 90%. The indirect ELISAs were further applied to test 595 goose serum samples. The results showed that the detection rates were 33.3% and 71.4% in GAstV-1-ELISA and GAstV-2-Cap-ELISA, respectively, and the co-detection rate was 31.1%, which indicates that the seroprevalence rate of GAstv-2 was higher than that of GastV-1, and the co-infection existed between GAstV-1 and GAstV-2. In summary, the developed GAstV-1-ELISA and GAstV-2-Cap-ELISA have high specificity, sensitivity, and reproducibility and can be used in the clinical detection of the antibody against GAstV-1 and GAstV-2.

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