不同抗原对猪瘟病毒抗体ELISA检测效果的影响

发布时间：2018-06-15 03:08

本文选题：ELISA + 检测效果　；参考：《中国兽药杂志》2017年06期

【摘要】：为研究不同抗原对猪瘟病毒抗体ELISA检测效果的影响,通过原核表达获取猪瘟病毒重组E2蛋白、E0蛋白、C蛋白和NS5B蛋白,大小分别约为35、42、16和80 kD。经Ni-NTA亲和层析柱纯化并利用Band Scan软件进行计算,纯度均在90%以上,满足ELISA检测包被用原料纯度的要求。将上述4种蛋白作为包被抗原进行ELISA,以10份企业阳性质控品血清和10份企业阴性质控品血清为检验指标比较不同抗原对猪瘟病毒抗体ELISA检测效果的影响。结果以E2蛋白为原料的包被板检测质控血清时,灵敏度和特异性均达到80%以上,能够满足猪瘟病毒抗体ELISA检测的要求,故选择E2蛋白作为包被抗原并进行相关检测试剂的研制。用研制的试剂与国际知名度高、产品质量好的美国IDEXX同类试剂对432份临床猪血清进行符合性检测,结果与美国IDEXX试剂的阳性符合率为95.53%,阴性符合率为86.56%,总符合率为91.67%,两种试剂检测结果具有较高的一致性。试验表明,用E2蛋白为包被抗原对猪瘟病毒抗体进行ELISA检测的效果最好,制备的检测试剂可用于临床猪瘟病毒血清抗体的检测,为今后猪瘟病毒抗体ELISA检测试剂的进一步研制奠定了基础。
[Abstract]:In order to study the effect of different antigens on the detection effect of swine fever virus antibody ELISA, recombinant E2 protein, E0 protein, C protein and NS5B protein were obtained by prokaryotic expression. The size was purified by Ni-NTA affinity chromatography column of 35,42,16 and 80 kD., respectively, and the purity was above 90%, which met the ELISA detection package. With the requirements of the purity of the raw materials, the above 4 proteins were used as the envelope antigen to be ELISA, the positive quality control serum of 10 enterprises and the serum of 10 negative quality control products were used as the test indexes to compare the effect of different antigens on the detection effect of swine fever virus antibody ELISA. The specificity is above 80%, and can meet the requirements of the detection of the antibody ELISA of the swine fever virus. So E2 protein is selected as the envelope antigen and the related detection reagents are developed. The developed reagents and the United States IDEXX reagents with high international popularity and good product quality test the conformance of 432 clinical pig serum, and the result is with the American IDEXX. The positive coincidence rate of the reagent was 95.53%, the negative coincidence rate was 86.56%, the total coincidence rate was 91.67%. The test results of the two reagents had high consistency. The test showed that the E2 protein as the envelope antigen had the best effect on the detection of swine fever virus antibody by ELISA, and the prepared detection reagent could be used for the detection of serum antibody in the clinical swine fever virus. The further development of ELISA detection reagent for swine fever virus antibody laid the foundation.
【作者单位】：郑州中道生物技术有限公司;河南中标检测服务有限公司;郑州大学生命科学学院;
【分类号】：S858.28

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