摘要
为建立便捷、快速检测猪圆环病毒2型(PCV2)抗体的免疫层析方法,本研究利用G蛋白标记胶体金制备金标垫,以PCV2病毒样颗粒(VLPs)和兔Ig G分别作为检测线与质控线组装胶体金试纸条。检测结果显示,该试纸条与A型塞内卡病毒(SVA)、猪细小病毒(PPV)、伪狂犬病毒(PRV)等猪其它病毒抗体阳性血清无交叉反应,仅对PCV2抗体阳性血清具有高度的特异性;对PCV 2阳性血清最低检测限为1∶6 400稀释度,敏感性较高;批内、批间变异系数CV均小于5%,重复性较好。试纸条在4℃保存6个月,其特异性和敏感性无明显变化。对采集的100份临床血清样品进行检测,该方法与标准ELISA的总符合率为98%。表明本研究建立的PCV2抗体的胶体金免疫层析检测方法具有敏感、特异、稳定等特点,可以做为评价PCV2疫苗免疫效果及其感染筛查的检测方法。
In order to establish a convenient and rapid immunochromatography method for the detection of porcine circovirus type 2(PCV2)antibody,in this study,protein G labeled colloidal gold was used to prepare gold-labeled mats,and PCV 2 virus-like particles(VLPs)and rabbit IgG were used as test line and control line respectively to assemble colloidal gold test strips.The test results showed that the test strip was highly sensitive as the PCV 2 antibody positive serum diluted to 1:6,400 can be detected;this text strip showed high specificity that was specific detecting PCV 2 antibody-positive serum but had no cross-reactivity with SVA,PPV,PRV as well as other viral positive sera of pigs.The inter-and intra-batch of coefficient of variation(CV)were less than 5%,showing high repeatability.The test strips were stored at 4℃for 6 months with no significant changes in specificity and sensitivity.100 clinical sera samples were tested and the total coincidence rate was 98%in comparison with the standard ELISA method.In conclusion,our results show that the PCV 2 antibody colloidal gold immunochromatographic method established in this study is sensitive,specific and stable,and can be used as a detection method for evaluating the immune effect of PCV 2 vaccine and viral infection screening.
作者
张禹
张韵
孙世琪
马小军
张小丽
郭慧琛
ZHANG Yu;ZHANG Yun;SUN Shi-qi;MA Xiao-jun;ZHANG Xiao-li;GUO Hui-chen(College of Veterinary Medicine,Gansu Agriculture University,Lanzhou 730070,China;State Key Laboratory of Veterinary Etiological Biology/National Foot-and-Mouth Disease Reference Laboratory/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2019年第7期705-709,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
国家重点研发计划(2017YFD0501100、2016YFD0500700)
