Computationally-driven epitope identification of PEDV N-protein and its application in development of immunoassay for PEDV detection
文献类型： 外文期刊
作者： Pang, Junzeng 1 ; Tian, Xiangqin 2 ; Han, Xiao 1 ; Yuan, Jiakang 1 ; Li, Linyue 1 ; You, Yonghe 3 ; Zhou, Yanlin 3 ; Xing, Guangxu 4 ; Li, Renfeng 1 ; Wang, Ziliang 1 ;
作者机构： 1.Henan Inst Sci & Technol, Coll Anim Sci & Vet Med, Xinxiang 453003, Peoples R China
2.Xinxiang Med Univ, Henan Key Lab Med Tissue Regenerat, Xinxiang 453003, Peoples R China
3.Xinxiang Med Univ, Sanquan Coll, Xinxiang 453000, Peoples R China
4.Henan Acad Agr Sci, Key Lab Anim Immunol, Minist Agr, Henan Prov Key Lab Anim Immunol, Zhengzhou 450002, Peoples R China
关键词： Porcine epidemic diarrhea virus; Epitopes of N protein; Monoclonal antibody; Quantum dots; Immunochromatographic assay
期刊名称：JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS （ 影响因子：3.4； 五年影响因子：3.3 ）
ISSN： 0731-7085
年卷期： 2023 年 235 卷
页码：
收录情况： SCI
摘要： The nucleocapsid (N) protein is a suitable candidate for early diagnosis of porcine epidemic diarrhea virus (PEDV). Here, we identified the linear B-cell epitopes of the PEDV N-protein by integrating a computationalexperimental framework and constructed three-dimensional (3D) structure model of the N protein using the ColabFold program in Google Colaboratory. Furthermore, we prepared the monoclonal antibodies against the predicted epitopes and recombinant N protein, respectively, and selected pairing mAbs (named 9C4 and 3C5) to develop a double-antibody sandwich immunochromatographic test strip using CdSe/ZnS quantum dots (QDs)labelled 9C4 and 3C5 as capture and detection antibodies, respectively. This strip can specifically detect PEDV within 10 min with a detection limit of less than 6.25 x 103 TCID50/mL. In comparison with RT-PCR for testing 90 clinical samples, the relative sensitivity and specificity of the strip were found to be 98.0% and 100%, respectively, with a concordance rate of 98.9% and a kappa value of 0.978, indicating that QDs-ICTS is a reliable method for the application of PEDV detection in clinical samples.
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