Development and evaluation of enzyme-linked immunosorbent assay based on recombinant nucleocapsid protein for detection of porcine epidemic diarrhea (PEDV) antibodies.
Development and evaluation of enzyme-linked immunosorbent assay based on recombinant nucleocapsid protein for detection of porcine epidemic diarrhea (PEDV) antibodies.
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DOI：
10.1016/j.vetmic.2007.02.014
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发表时间：
2007-07-20
影响因子：
3.3
通讯作者：
Liu J
中科院分区：
文献类型：
--
作者：
Hou XL;Yu LY;Liu J
以重组大肠杆菌核衣壳蛋白（N蛋白）为抗原，建立了诊断猪流行性腹泻（PEDV）的酶联免疫吸附试验（ELISA），并对其敏感性和特异性进行了评价。克隆了编码N蛋白的N基因，并在E.杆菌重组N蛋白迁移至48 kDa处，与6个组氨酸标签特异性单克隆抗体进行免疫印迹反应。重组N蛋白ELISA（rnELISA）在（80）个无PEDV的个体中显示98.7%的特异性，并且在（103）个有PEDV的临床样品中显示98%的灵敏度。在检测884份现场样本时，SN和rnELISA之间的总体一致率为88.3%。这些结果表明，核衣壳蛋白可能是一个有用的抗原，用于血清诊断PEDV，它也表明，ELISA是一种高度敏感和特异性的检测抗体PEDV。
An enzyme-linked immunosorbent assays (ELISA) based on recombinant nucleocapsid (N) protein generated in Escherichia coli was evaluated for its sensitivity and specificity for diagnosis of porcine epidemic diarrhea (PEDV) infection. The N gene encoding the N protein was cloned and expressed as a fusion protein with His tag protein in E. coli. The recombinant N protein was migrated at 48 kDa and reacted with six histidine tag specific monoclonal antibody by immunoblotting. Recombinant N protein ELISA (rnELISA) demonstrated 98.7% specificities among (80) PEDV-free individuals, and 98% sensitivity ranging among (103) clinical samples with PEDV. On testing 884 field samples, an overall agreement of 88.3% was generated between the SN and rnELISA. Taken together, these results indicated that nucleocapsid protein may be a useful antigen for the sera-diagnosis of PEDV and it was also suggested that the ELISA is a highly sensitive and specific test for detecting antibodies to PEDV.
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