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Position: Home > Articles > Establishment of an Immunofluorescence Assay for Direct Detection of Avian Reovirus China Animal Health Inspection 2012 (11) 35-39

直接免疫荧光检测禽呼肠孤病毒方法的建立

作  者:
谢志勤;谢芝勋;刘加波;庞耀珊;邓显文;谢丽基;范晴
单  位:
广西壮族自治区兽医研究所
关键词:
禽呼肠孤病毒;结构蛋白σ3;直接免疫荧光
摘  要:
本研究利用经纯化的原核表达禽呼肠孤病毒(ARV)结构蛋白σ3作为免疫抗原,免疫兔子三次,血清效价达到27时采血分离血清。用硫酸铵沉淀法浓缩纯化血清中的IgG,纯化的IgG用荧光素FITC加以标记。利用制备的荧光标记特异性抗体和对反应条件进行优化,建立了表达σ3蛋白荧光抗体检测ARV的诊断方法。结果经纯化获得特异性抗ARV结构蛋白σ3的荧光标记IgG抗体。用标记的抗体检测ARV病毒感染的单层细胞,出现特异性的荧光。用建立的σ3-FA方法检测接种疑似ARV感染的病料的鸡胚原代成纤维细胞,检出的阳性样品与病毒分离结果一致。表明本研究建立的σ3-FA方法检测ARV病毒有较好的特异性,可用于ARV感染的临床样品检测。
译  名:
Establishment of an Immunofluorescence Assay for Direct Detection of Avian Reovirus
作  者:
Xie Zhiqin,Xie Zhixun,Liu Jiabo,Pang Yaoshan,Deng Xianwen,Xie Liji,Fan Qing(Guangxi Veterinary Research Institute,Guangxi Key Laboratory of Animal Vaccine and New Technology,Nanning, Guangxi, 530001,China)
关键词:
avian reovirus;σ3 protein;direct immunofluorescence
摘  要:
The purified prokaryotic expression σ3 structure protein of avian reovirus (ARV) as the immunogen was injected into rabbit.The serum was isolated from the blood when the antibody titer was over 2~7.The immunoglobulin (IgG) from the serum was precipitated by ammonium sulfate and labeled with fluorescein isothiocyanate (FITC).A direct immunofluoresence assay for detecting avian reovirus was established by optimizing the reaction conditions.Then the clinical samples were tested.The results showed that the purification of IgG labeled with FITC was got.The immunofluoresence from positive sample infected by reovirus S1733 strain was shown under the microcopy.There was no cross with other viruses.It had the same result as the virus isolation.The method could be used for detection of ARV from the clinically infected samples in the future.

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