文献类型： 外文期刊

第一作者： Han, Weiguo

作者： Han, Weiguo;Ma, Zhiqian;Li, Zhiwei;Chang, Chuanzhe;Yuan, Yue;Li, Yongqi;Feng, Ran;Zheng, Congsen;Shi, Zhengwang;Tian, Hong;Zheng, Haixue;Xiao, Shuqi

作者机构：

关键词： PEDV; Nucleocapsid protein; Monoclonal antibodies; DAS-qELISA; Antigen detection

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.3； 五年影响因子：5.1 ）

ISSN： 0175-7598

年卷期： 2024 年 108 卷 1 期

页码：

收录情况： SCI

摘要： Porcine epidemic diarrhea (PED), a contagious intestinal disease caused by the porcine epidemic diarrhea virus (PEDV), has caused significant economic losses to the global pig farming industry due to its rapid course and spread and its high mortality among piglets. In this study, we prepared rabbit polyclonal antibody and monoclonal antibody 6C12 against the PEDV nucleocapsid (N) protein using the conserved and antigenic PEDV N protein as an immunogen. A double-antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-qELISA) was established to detect PEDV using rabbit polyclonal antibodies as capture antibodies and horseradish peroxidase (HRP)-labeled 6C12 as the detection antibody. Using DAS-qELISA, recombinant PEDV N protein, and virus titer detection limits were approximately 0.05 ng/mL and 103.02 50% tissue culture infective dose per mL (TCID50/mL), respectively. There was no cross-reactivity with porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (PoRV), porcine pseudorabies virus (PRV), porcine deltacoronavirus (PDCoV), or porcine circovirus (PCV). The reproducibility of DAS-qELISA was verified, and the coefficient of variation (CV) for intra- and inter-batch replicates was less than 10%, indicating good reproducibility. When testing anal swab samples from PEDV-infected piglets using DAS-qELISA, the coincidence rate was 92.55% with a kappa value of 0.85 when using reverse transcription-polymerase chain reaction (RT-PCR) and 94.29% with a kappa value of 0.88 when using PEDV antigen detection test strips, demonstrating the reliability of the method. These findings provide fundamental material support for both fundamental and practical studies on PEDV and offer a crucial diagnostic tool for clinical applications.Key points center dot A new anti-PEDV N protein monoclonal antibody strain was prepared center dot Establishment of a more sensitive double antibody sandwich quantitative ELISA center dot DAS-qELISA was found to be useful for controlling the PEDV spreadKey points center dot A new anti-PEDV N protein monoclonal antibody strain was prepared center dot Establishment of a more sensitive double antibody sandwich quantitative ELISA center dot DAS-qELISA was found to be useful for controlling the PEDV spreadKey points center dot A new anti-PEDV N protein monoclonal antibody strain was prepared center dot Establishment of a more sensitive double antibody sandwich quantitative ELISA center dot DAS-qELISA was found to be useful for controlling the PEDV spread

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