摘要
为建立快速定量检测家禽样本中禽白血病病毒抗原的方法,对禽白血病病毒p27蛋白单克隆抗体细胞株2E5和3D5进行复苏、鉴定,通过反应条件优化,建立了禽白血病病毒抗原高敏荧光微球快速检测方法。该方法能够快速定量检测家禽样本中禽白血病病毒,敏感性高、特异性强,对其他家禽病原无交叉反应且具有良好的检测重复性。对采自全国的240份临床家禽样品进行检测,同时用美国IDEXX公司生产的ELISA抗原试剂盒开展比较,结果发现两种方法符合率达93.98%,其中阳性样品符合率为90.83%,阴性样品符合率为95.83%;组内与组间变异系数分别低于10%和15%。本研究建立的禽白血病病毒p27抗原高敏荧光微球快速检测方法,特异性高、敏感性强,操作简单、快速,具有较高的应用推广价值。
In order to establish a rapid and quantitative method for detection of avian leukosis virus(ALV)in poultry samples,2E5 and 3D5,the monoclonal antibody cell strains of ALV p27 protein were revived and identified,a highly sensitive and rapid fluorescence quantitative detection method,with high sensitivity and specificity and good test repeatability and no cross reaction to other pathogens,was established through optimization of reaction conditions,by which,ALV could be rapidly and quantitatively detected from poultry samples.240 clinical samples collected across China were detected and compared with ELISA kit produced by IDEXX.It was found that the coincidence rate of the two methods was 93.98%,in which the coincidence rate of positive samples was 90.83%,and that of negative ones was 95.83%.The coefficients of variation in intra group and inter group were lower than 10%and 15%,respectively.In conclusion,the established method was worthy of being applied and expanded as supported by its high specificity and sensitivity,easy and rapid operation.
作者
孙雨
刘亚涛
王睿男
蒋菲
马英
李秀梅
王文
原小燕
顾小雪
王传彬
Sun Yu;Liu Yatao;Wang Ruinan;Jiang Fei;Ma Ying;Li Xiumei;Wang Wen;Yuan Xiaoyan;Gu Xiaoxue;Wang Chuanbin(China Animal Disease Control Center,Beijing 102600,China;Xinjiang Uygur Autonomous Region Center for Animal Disease Prevention and Control,Urumqi,Xinjiang 830000,China;Luoyang Institute of Modern Biological Technology Co.,Ltd.,Luoyang,Henan 471000,China)
出处
《中国动物检疫》
CAS
2021年第5期96-103,共8页
China Animal Health Inspection
基金
国家重点研发计划项目(2016YFD0500809)。
关键词
禽白血病
p27融合抗原
单克隆抗体
荧光微球
快速定量检测
avian leukemia
p27 fusion antigen
monoclonal antibody
fluorescent microsphere
rapid quantitative detection
