Development of a unique sandwich enzyme-linked immunosorbent assay based on monoclonal antibodies for the specific detection of the egg drop syndrome virus

文献类型： 外文期刊

第一作者： Wei, Qiang

作者： Wei, Qiang;Liu, Yunchao;Li, Qingmei;Jin, Qianyue;Chai, Shujun;Xing, Guangxu;Zhang, Gaiping;Gao, Yanling;Song, Yapeng;Zhang, Gaiping

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关键词： Egg drop syndrome virus; sandwich enzyme-linked immunosorbent assay; fibre protein; monoclonal antibody; detection of antigen; limit of detection

期刊名称：AVIAN PATHOLOGY （ 影响因子：2.8； 五年影响因子：3.2 ）

ISSN： 0307-9457

年卷期： 2024 年 53 卷 2 期

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收录情况： SCI

摘要： An abrupt decrease in egg production and quality in laying hens can be brought on by the avian adenovirus known as the egg drop syndrome virus (EDSV). So far, there are no efficient commercial diagnostic approaches for EDSV. In the present study, we first purified two monoclonal antibodies (mAbs 5G4 and 6G6) specifically against the EDSV fibre protein. The mAb 5G4 was then employed as capture antibody. Additionally, the detection antibody 6G6 (HRP-6G6) was conjugated with horseradish peroxidase. Consequently, based on these two mAbs, we developed a sandwich enzyme-linked immunosorbent assay (ELISA) for EDSV detection. Specificity evaluation demonstrated that only EDSV gave a positive result while other avian viruses showed negative results. Sensitivity investigation showed that the limit of detection of EDSV was 102.9 TCID50/ml and the limit of detection of the purified His-Fiber was 5 ng/ml. Furthermore, the screening of samples from infected chickens showed that this method was very effective and could be applied to the detection of EDSV in infected samples. In summary, the sandwich enzyme-linked immunosorbent assay developed here provides an efficient, sensitive, and large-scale method for the specific diagnosis of EDSV.RESEARCH HIGHLIGHTSA sandwich ELISA was developed to detect EDSV using the mAbs 5G4 and HRP-6G6.The sandwich ELISA maintained high specificity and sensitivity.The sandwich ELISA had equivalent consistency with real-time PCR assay.

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