数字农科院2.0

Development of a Blocking ELISA for Detection of Serum Neutralizing Antibodies Against Duck Adenovirus Type 3

文献类型： 外文期刊

作者： Mei Tang;Xiaona Shi;Yifei Xiong;Chunxiu Yuan;Qinmin Zhu;Minfan Huang;Bangfeng Xu;Qinfang Liu;Xue Pan;Zhifei Zhang;Qiaoyang Teng;Minghao Yan;Dawei Yan;Zejun Li

作者机构：

关键词： blocking ELISA;detection;duck adenovirus type 3 (DAdV-3);monoclonal antibody

期刊名称： Microorganisms

ISSN： 2076-2607

年卷期： 2025 年 13 卷 11 期

页码：

收录情况： SCIE（2025版）

摘要： In 2014, Duck Adenovirus type 3 (DAdV-3) emerged in Muscovy ducks and has since spread rapidly across China, causing significant economic losses to the duck industry. Given this situation, the development of reliable diagnostic tools is crucial for effective disease control. In this study, a neutralizing monoclonal antibody (mAb) 2F12 specific to DAdV-3 was generated, which showed a blocking rate of over 70% and a neutralization titer of up to 1:794. A blocking enzyme-linked immunosorbent assay (b-ELISA) was further developed based on mAb 2F12 to efficiently detect neutralizing antibodies against DAdV-3. The cut-off values of percent inhibition (PI) were set based on testing 84 negative duck serum samples, with a value below 16.79% (mean ((Formula presented.)) + 2 standard deviations (SD)) for negative sera and over 21.62% ((Formula presented.) + 3SD) for positive sera. The b-ELISA exhibited a high specificity, reacting exclusively with DAdV-3 positive serum and showing no cross-reactivity with other representative positive sera tested. Additionally, the b-ELISA showed significantly higher sensitivity than the serum neutralization test (SNT), detecting antibodies 16-fold greater than the endpoint dilution of the SNT. The established b-ELISA, validated with 90 field serum samples from six duck farms, was well-suited for clinical detection of DAdV-3 antibodies and for monitoring post-vaccination antibody levels, representing a significant advancement in DAdV-3 detection and prevention.

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