摘要
猪繁殖与呼吸综合征病毒(HP-PRRSV)能引起猪的繁殖障碍等疾病,是世界养猪业中的一大难题。本研究以HP-PRRSV病毒的N(核衣壳)蛋白为研究基础,原核表达N蛋白后,获得纯化蛋白,进行动物免疫,获得单克隆抗体株,优化间接ELISA和胶体金试纸条的条件。纯化后重组核衣壳蛋白的纯度大于90%;Western blot出现明显的反应条带;建立间接ELISA试验方法,且无明显交叉反应;筛选出9株针对N蛋白的抗体,特异性较好;组装的胶体金试纸条可检测最低稀释抗原质量浓度约为5mg/L。该试验为快速检测PRRSV、提高检测速率奠定一定的基础。
Porcine reproductive and respiratory syndrome virus(PRRSV)has become a problem to the swine industry in the world wide.PRRSV causes a persistent and sometimes severe disease which is characterized by respiratory problems,weight loss and poor growth performance.In this study,the N protein purified by the nickel agarose resin chromatographic analysis column,was used for an appraisal of Western blot and immunization.At last,we optimized conditions of the indirect ELISA and the colloidal gold immunochromatographic test strips.The results showed that the purity of goal protein reached as high as 90%,and N protein had the good immunogenicity and specificity;by the optimization of conditions,an indirect ELISA was established for detecting antibodies against N protein in swine serum with good sensitivity and specificity;9antibodies had no cross reaction with some viruses;The lowest detectable limit was N 5mg/L protein.The strip laid a fundation for rapid detection of PRRSV.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2014年第6期853-858,共6页
Chinese Journal of Veterinary Science
基金
河北省重大科技支撑计划资助项目(11230405D)
关键词
猪繁殖与呼吸综合征病毒
核衣壳蛋白
单克隆抗体
胶体金试纸
RRSV
nucleocapsid protein
monoclonal antibodies
colloidal gold immunochromatographic test strips
