猪圆环病毒2型合成肽间接ELISA抗体检测方法的建立和评估  

Establishment and Evaluation of a Synthetic Peptide-based Indirect ELISA for Detection of Antibodies against Porcine Circovirus Type 2

作　　者：栗利芳[1,2] 刘新月 张蕾 董春娜 肖进 汪明 郭鑫[1] LI Li-fang;LIU Xin-yue;ZHANG Lei;DONG Chun-na;XIAO Jin;WANG Ming;GUO Xin(College of Veterinary Medicine,China Agricultural University,Beijing 100193,China;Key Laboratory of Veterinary Bioproduction and Chemical Medicine of the Ministry of Agriculture,Engineering and Technology Research Center for Beijing Veterinary Peptide Vaccine Design and Preparation,Zhongmu Institutes of China Animal Husbandry Industry Co.Ltd,Beijing 100095,China)

机构地区：[1]中国农业大学动物医学院,北京海淀100193 [2]中牧实业股份有限公司,农业农村部兽用生物制品与化药重点实验室,北京市兽用多肽疫苗设计与制备工程技术中心,北京海淀100095

出　　处：《中国兽医杂志》2021年第9期16-21,26,共7页Chinese Journal of Veterinary Medicine

摘　　要：本试验根据猪圆环病毒2型(PCV2)Cap蛋白表位,结合文献报道及软件预测,设计合成肽抗原;再用PCV2阳性血清进行筛选,优选出4条肽的混合肽段作为包被抗原;继而通过最适条件摸索,建立PCV2合成肽间接ELISA抗体检测方法,并从该方法的敏感性、特异性、重复性及其他试剂盒进行比对试验等关键性指标进行评估。结果显示,间接ELISA的敏感性为95.0%,特异性为97.5%,批内变异系数在3.4%~7.2%,批间重复性变异系数在4.8%~8.6%。比对试验结果显示,本间接ELISA与PCV2 Cap蛋白包被试剂盒符合率为91.5%;与猪圆环病毒2-dCap-ELISA抗体检测试剂盒(锐捷)符合率为94.6%。因此,本试验建立的猪圆环合成肽间接ELISA抗体检测方法敏感性好、特异性强和重复性好,可用于临床PCV2血清抗体检测及作为疫苗免疫效果评价的技术手段。Four synthetic peptides were designed and optimized from published porcine circovirus type 2(PCV2)Cap proteins through software prediction,and synthesized by the solid phase method.An indirect ELISA based on these synthetic peptides was developed for detection of specific antibodies against PCV2 in pigs.The results showed that this indirect ELISA displayed no cross-reaction with other swine sera,and had a sensitivity of 95.0% and a specificity of 97.5%.The variant coefficient of this indirect ELISA within batch and between batches ranged from 3.4% to 7.2% and from 4.8% to 8.6%,respectively.This indirect ELISA had coincidence rates of 91.5% and 94.6% with Cap-ELISA method and PCV2-dCap-ELISA Kit,respectively.These data suggested that the indirect ELISA is cost-effective,sensitive,specific,and repeatable,and can be applied for clinical detection of serum antibodies against PCV2 in pigs and also as a technical means for evaluating vaccine effectiveness.

关 键 词：猪圆环病毒2型 合成肽 间接ELISA 抗体检测 

分 类 号：S852.651[农业科学—基础兽医学]