| 鸭疫里默杆菌单克隆抗体的研制及免疫胶体金检测方法的建立 |
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| 引用本文： | 陈素娟,陈冰,孙化露,高以明,彭大新,刘秀梵.鸭疫里默杆菌单克隆抗体的研制及免疫胶体金检测方法的建立[J].兽医大学学报,2012(11):1651-1655. |
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| 作者姓名： | 陈素娟 陈冰 孙化露 高以明 彭大新 刘秀梵 |
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| 作者单位： | [1]扬州大学兽医学院,农业部畜禽传染病重点开放实验室,禽类预防医学教育部重点实验室,江苏扬州225009 [2]南京出入境检验检疫局,南京210001 |
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| 基金项目： | 公益性行业（农业）科研专项资助项目（201003012）;江苏省检验检疫局科研资助项目（2009KJ04）;教育部创新团队资助项目（IRT0978）;青蓝工程[苏教师（2010）27号];禽类预防医学教育部重点实验室和教育部创新团队资助项目（IRT0978） |
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| 摘 要： | 为快速检测临床样品中鸭疫里默氏杆菌（RA），建立检测RA的免疫胶体金方法。以血清1型和2型RA分别免疫BALB／c小鼠，通过细胞融合技术制备单克隆抗体。经ELISA筛选获得4株单克隆抗体2F7、3G9、5G7和2E6，5G7可与1、2、3、11型RA交叉反应，而其他3株与1、2、3型RA有交叉反应。选取单克隆抗体5G7进行纯化和胶体金标记，制备免疫胶体金试纸条，可检测1、2、3、11型RA，而与禽巴氏杆菌、大肠杆菌、沙门菌无反应。对RA模拟样品最低检出剂量为6×10^3 CFU细菌，与细菌分离方法相比，免疫胶体金试纸条对临床病料中RA检测的敏感性为100％，符合率为88．9％。
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| 关 键 词： | 鸭疫里默杆菌 单克隆抗体 免疫胶体金 |
Development of monoclonal antibodies and gold immunochromatographic assay for detecting Riemerella anatipesti f er |
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| Authors: | CHEN Su-juan CHEN Bing SUN Hua-lu PENG Da-xin GAO Yi-ming LIU Xiu-fan |
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| Institution: | 1. Key Laboratory of Animal Infectious Disease Ministry of Agriculture, College of Veterinary Medicine Yangzhou University ,Yangzhou ,J iangsu 225009 ,China ; 2. Nanjing Entry-Exit Inspec- tion and Quarantine Bureau, Nanjing 210001 ,China) |
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| Abstract: | To develop a rapid and accurate test based on the gold immunochromatographic assay （GICA） for the detection of Riemerella anatipestifer （RA）, BALB/c mice were immunized with whole cells of serotype 1 and 2 RA strains,respectively. The spleen cells of immunized mouse were fused with myeloma cells（SP2/0）. Four positive hybridomas cell strains were identified and named as 2F7,3G9,5G7 and 2E6, respectively. Monoclonal antibodies （mAb） 5G7 had cross reaction with serotype 1,2,3 and 11 strains,while the other 3 mAbs with serotype 1,2 and 3 strains. GICA was established by purified and gold labeled mAb 5G7. It could detect all four serotype strains of RA, but not Pasteurella multocida ,E. coli and salmonella. The minimum detectable limit of GICA for RA mimic clinical samples was6×10^3 CFU. The detection results for clinical samples showed that the sensitivity and coincidence rate of GICA were 100% and 88. 9%, respectively, when it was compared with bacterial isolation method. |
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| Keywords: | Riemerella anatipestifer monoclonal antibody lmmune colloidal gold |
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