猪圆环病毒2型Cap蛋白琼脂扩散试验检测方法的建立  

Establishment of an Agar-gel Precipitation Test for PCV2 Cap Protein

作　　者：庄金秋[1,2] 梅建国 张颖[1] 董林 李峰[1] Zhuang Jinqiu;Mei Jianguo;Zhang Ying;Dong Lin;Li Feng(Shandong Binzhou Animal Science&Veterinary Medicine Academy,Binzhou,Shangdong 256600,China;Binzhou Bio-carrier Biotechnology Co.,Ltd.,Binzhou,Shangdong 256600,China)

机构地区：[1]山东省滨州畜牧兽医研究院,山东滨州256600 [2]滨州贝尔凯瑞生物技术有限公司,山东滨州256600

出　　处：《中国动物检疫》2020年第9期113-117,共5页China Animal Health Inspection

基　　金：山东省自然科学基金资助项目(ZR2017MC056);山东省现代农业产业技术体系生猪产业创新团队项目(SDAIT-08-17)。

摘　　要：为便于快速、准确检测猪圆环病毒2型(PCV2),减少其对我国养猪业的威胁,建立了PCV2 Cap蛋白琼脂扩散试验(AGP)检测方法。应用表达制备好的PCV2 Cap蛋白,与弗氏佐剂乳化后免疫家兔,制备多克隆抗体并进行抗体效价测定。确定AGP最佳工作条件,并进行抗体特异性、稳定性及符合率试验。结果显示:制备的多克隆抗体ELISA效价可达1:12800,AGP效价可达1:32,表明其具有良好的特异性、稳定性;对20份PCV2 Cap蛋白抗原进行检测,发现其结果与应用标准猪血清抗体检测结果符合率为100%。结果表明,制备的抗PCV2 Cap蛋白多克隆抗体可替代标准抗体,用于PCV2 Cap蛋白的AGP试验检测。本研究为PCV2检测以及后续的免疫学特性研究提供了技术支撑。In order to rapidly and accurately detect porcine circovirus type 2(PCV2)so as to reduce the threat to pig industry in China,an agar-gel precipitation(AGP)test was established for detection of PCV2 Cap protein.Some rabbits were vaccinated by emulsifying PCV2 Cap protein with Freund's adjuvant to prepare polyclonal antibodies and the titer was determined,then the specificity,stability and coincidence rate of the antibody were tested after the optimal conditions of AGP test were determined.The results showed that the titer of the polyclonal antibodies could reach up to 1:12800 by ELISA,and AGP titer was up to 1:32,showing good specificity and stability;20 samples of PCV2 Cap protein were tested,and the results completely conformed to those by detection of standard pig serum antibodies.Therefore,the polyclonal antibodies against PCV2 Cap protein could replace the standard antibodies,and be used for AGP test.In conclusion,some support was provided for PCV2 detection and subsequent study on the immunological characteristics.

关 键 词：PCV2 Cap蛋白 多克隆抗体 琼脂扩散试验 

分 类 号：S852.65[农业科学—基础兽医学]