| 牛轮状病毒双抗体夹心ELISA检测方法的建立 |
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| 引用本文： | 侯美如,侯喜林,高俊峰,刘振格,王杰,杨明发. 牛轮状病毒双抗体夹心ELISA检测方法的建立[J]. 黑龙江八一农垦大学学报, 2011, 23(3): 19-23 |
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| 作者姓名： | 侯美如 侯喜林 高俊峰 刘振格 王杰 杨明发 |
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| 作者单位： | 黑龙江八一农垦大学动物科技学院,大庆,163319 |
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| 基金项目： | 黑龙江农垦总局攻关课题 |
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| 摘 要： | 用差速离心法纯化的牛轮状病毒(BRV)分别免疫鼠和兔,制备了鼠抗BRV和兔抗BRV超免疫血清,并用层析方法进行了纯化,建立了检测BRV的双抗体夹心ELISA方法.该双抗体夹心ELISA的最佳反应条件为:兔抗BRV IgG包被浓度为10μg·mL-1,鼠抗BRV IgG工作浓度为5 μg·mL-1,样品反应时间60 mi...
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| 关 键 词： | 牛轮状病毒 双抗体夹心酶联免疫吸附试验 检测 |
Development of a Double Antibody Sandwich ELISA for Detection of Bovine Rotavirus |
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| Hou Meiru,Hou Xilin,Gao Junfeng,Liu Zhenge,Wang Jie,Yang Mingfa. Development of a Double Antibody Sandwich ELISA for Detection of Bovine Rotavirus[J]. journal of heilongjiang bayi agricultural university, 2011, 23(3): 19-23 |
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| Authors: | Hou Meiru Hou Xilin Gao Junfeng Liu Zhenge Wang Jie Yang Mingfa |
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| Affiliation: | (College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Daqing 163319) |
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| Abstract: | Two kinds of hyperimmune sera,mouse-anti-rotavirus IgG and rabbit-anti-rotavirus IgG,were prepared by inoculating respectively piglets and rabbits with bovine rotavirus(BRV) purified by differential centrifugation,and purified by affinity chromatography.A double antibody sandwich ELISA for detection of BRV was developed based on the two kinds of IgG.The optimal coating concent ration of rabbit-anti-rotavirus IgG was 10 μg·mL-1,the optimal working concent ration of mouse-anti-rotavirus IgG was 5 μg·mL-1,the reaction time of sample was 60 min,and the optimal working dilution of HRP-labelled goat-anti-mouse IgG was 1:5 000.The positive standard value was 0.432(OD450 nm).The coefficient of variation of reproducibility was less than 10%,and at least 1.41 μg·mL-1 antigen could be detectable.The ELISA had no cross-reaction with IBRV,BCV,BRSV.Forty clinical fecal samples were detected by the ELISA and RT-PCR with agreement rate of 95%.The results revealed that the ELISA possessed good specificity and reproducibility,and higher sensitivity,indicating a suitable method for rapid detection of BRV. |
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| Keywords: | Bovine rotavirus double antibody sandwich ELISA detection |
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