Development of sandwich Enzyme-Linked Immunosorbent Assay for the detection of porcine epidemic diarrhea virus in fecal samples
文献类型： 外文期刊
作者： Gui, Rui 1 ; Shi, Hong-yan 2 ; Liu, Wei 1 ; Feng, Li 2 ; Yang, Ke-li 1 ; Guo, Rui 1 ; Liang, Wan 1 ; Yuan, Fang-yan 1 ; Duan 1 ;
作者机构： 1.Hubei Acad Agr Sci, Anim Husb & Vet Inst, Key Lab Prevent & Control Agents Anim Bacteriosis, Minist Agr, Wuhan 430064, Hubei, Peoples R China
2.Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol, Harbin 150001, Heilongjiang, Peoples R China
3.Islamia Univ Bahawalpur, Univ Coll Vet & Anim Sci, Bahawalpur 63100, Pakistan
关键词： PEDV; S1; MAb; S-ELISA
期刊名称：MICROBIAL PATHOGENESIS （ 影响因子：3.738； 五年影响因子：3.663 ）
ISSN： 0882-4010
年卷期： 2018 年 122 卷
页码：
收录情况： SCI
摘要： Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in new-born piglets with subsequent economic losses to swine industry. In the current study, gene encoding of 381aa-792aa spike protein (S1) with the main epitope relative to virus neutralization of PEDV was amplified by RT-PCR and inserted into vector pET-30A(+). The plasmid was transferred into Escherichia coli BL21 (DE3). Meanwhile, recombinant protein expression was induced by isopropyl-P-galactopyranoside (IPTG). After denaturation and renaturation of inclusion bodies, the Si protein was obtained by using purified recombinant S1 protein in immunized female BALB/c mice. Monoclonal antibodies (MAb) against Si protein, named 4C7 by hybridoma technique were gained successfully. The result showed that MAb can specifically respond to S1 protein and PEDV via ELISA, Western bolt and immunofluorescence assay methods. A sandwich ELISA (S-ELISA) was established by using the captured monoclonal antibodies 4C7. The sensitivity and specificity were compared between S-ELISA and RTPCR, which showed similar sensitivity and specificity. This work indicated that S-ELISA would be a significant tool alongside a specific diagnostic reagent for PEDV in future.
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