Li R, Tian X, Yu Y, Qiao S, Wang Z, Ma J
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang 453003, China.
Key Laboratory of Animal Immunology of the Ministry of Agriculture, Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, China.
Pol J Vet Sci. 2018 Mar;21(1):139-147. doi: 10.24425/119032.
An immunochromatographic test strip was developed for the detection of antibodies against porcine epidemic diarrhea virus (PEDV) in porcine serum. In this test, the recombinant nucleocapsid protein of PEDV was expressed in soluble form and purified by using Ni-NTA resin and gel filtration chromatography. The purified recombinant N protein conjugated with colloidal gold was dispensed on a conjugate pad as the detector. Staphylococcal protein A and rabbit anti-N protein IgG were blotted on a nitrocellulose membrane for the test and control lines, respectively. The immunochromatographic test strip specifically detected PEDV antibodies within 10 min and had higher sensitivity (96.0%) and specificity (90.8%) than those of commercial enzyme-linked immunosorbent assay (ELISA) kits. Our newly developed strip has great potential for the early diagnosis of PEDV infection.
开发了一种免疫层析试纸条,用于检测猪血清中抗猪流行性腹泻病毒(PEDV)的抗体。在该检测中,PEDV的重组核衣壳蛋白以可溶形式表达,并通过使用镍-氮川三乙酸(Ni-NTA)树脂和凝胶过滤色谱法进行纯化。与胶体金结合的纯化重组N蛋白作为检测剂点样在结合垫上。葡萄球菌蛋白A和兔抗N蛋白IgG分别印迹在硝酸纤维素膜上作为检测线和对照线。该免疫层析试纸条可在10分钟内特异性检测PEDV抗体,并且比市售酶联免疫吸附测定(ELISA)试剂盒具有更高的灵敏度(96.0%)和特异性(90.8%)。我们新开发的试纸条在PEDV感染的早期诊断方面具有巨大潜力。