新城疫病毒液相芯片技术检测方法的建立  

Development of the detection method of Newcastle disease virus utilizing liquid chip technology

作　　者：朱余军[1] 丛锋[1] 饶丹[1] 伍妙梨[1] 肖丽 练月晓 黄碧洪 黄韧[1] 郭鹏举[1] 陈梅丽[1] 

机构地区：[1]广东省实验动物监测所广东省实验动物重点实验室,广东广州510663

出　　处：《中国兽医科学》2017年第10期1264-1268,共5页Chinese Veterinary Science

基　　金：广东省科技计划项目(2015B070701003;2016A040403065)

摘　　要：为建立一种新城疫病毒的双抗夹心液相芯片技术检测方法,将捕获抗体偶联到67号微球,对偶联抗体量、检测抗体和SA-PE抗体的工作浓度进行确定,用建立的液相芯片方法进行特异性、灵敏度、重复性及临床样品的检测。结果显示,1×10~6个微球的偶联抗体量为25 g,最优的检测抗体与SA-PE的工作浓度分别为2 g/mL和4 g/mL。该方法对其他病原无交叉反应,具有较好的特异性;检测的敏感效价为0.0625 HA,比传统的HA灵敏度高;批内、批间的变异系数分别为2.0%和6.7%;对60份临床样品检出率为5%,与PCR的检出率100%相符。结果表明,本试验建立的新城疫液相芯片检测方法具有特异性好、灵敏度高、重复性好等特点,为新城疫病毒的检测提供了一种新方法。To develop a detection method of Newcastle disease virus utilizing double-antibody sandwich liquid chip technology,the capture antibodies were conjugated to the 67 microshpere.The coupling capacity of capture antibodies,the working concentration of detection antibody and SA-PE were optimized.At the same time the specificity,sensitivity,stability and clinical samples of the method were analyzed.The optimal coupling capacity of capture antibody was 25 g of 1×10~6 microspheres,and the optimal working concentration of detection antibody and SA-PE was 2 g/mL and 4 g/mL,respectively.The results showed that the method had not cross reactions with other viruses.The detection limit reached 0.0625 HA,and the sensitivity was higher than HA.Reproducibility analysis showed that the intra-and inter-assay coefficient of variation was 2.0% and 6.7%,respectively.The detection rate of60 clinical samples was 5%.Compared to PCR,there had a same detection rate.These results indicated that the method was high specificity,sensitivity and stability.It is a new method for detectiong Newcastle disease virus.

关 键 词：新城疫病毒 液相芯片 检测方法 

分 类 号：S852.659.5[农业科学—基础兽医学]