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大菱鲆疾病早期快速检测方法——胶体金免疫层析试纸的研制与建立
引用本文:王蔚芳,柴书军,刘庆堂,雷霁霖,丁福红,洪磊,刘新富,苏柯.大菱鲆疾病早期快速检测方法——胶体金免疫层析试纸的研制与建立[J].中国工程科学,2012,14(2):8-13.
作者姓名:王蔚芳  柴书军  刘庆堂  雷霁霖  丁福红  洪磊  刘新富  苏柯
作者单位:1. 中国水产科学研究院黄海水产研究所,青岛市海水鱼类种子工程与生物技术重点实验室 青岛,266071
2. 河南省农业科学院,河南省动物免疫学重点实验室,郑州450002
3. 青岛通用水产养殖有限公司 青岛266404
基金项目:国家鲆鲽类产业技术体系(CARS-50);中国博士后科学基金(20100471499);山东省博士后创新项目专项资金(200902008); 中国水产科学研究院黄海水产研究所基本科研业务费项目(20603022011018)
摘    要:使用成分单一的牛血清白蛋白(BSA)为模拟病原,以胶体金标记兔抗血清(即大菱鲆免疫球蛋白多抗)作为检测示踪物,并分别将BSA和葡萄球菌A蛋白印记到硝酸纤维素膜上制成检测线和对照线,通过一系列工艺创制与组装配套,首次成功制备了一套完整的大菱鲆抗体快速检测试纸。采用大菱鲆抗BSA血清作为阳性样本,以健康大菱鲆血清作为阴性样本,用以检验试纸的性能,并与酶联免疫吸附实验(ELISA)法检测结果相比较。结果表明:本试纸检测抗体的特异性与敏感性均很高,与ELISA方法相当,而且使用方便,不需专业技能和额外的试剂与辅助仪器设备,5 min内即可用裸眼获得观察结果,很适合于基层生产操作及户外调研使用。以该实验为基础建立起来的抗体检测试纸,亦可推广应用于其他病害抗体的检测,可为鱼类疾病早期发生提供简易、快捷和操作性强的诊断方法。

关 键 词:大菱鲆  抗体  试纸  现场快速检测  胶体金免疫层析  疾病
收稿时间:2011/10/13 0:00:00
修稿时间:2011/10/13 0:00:00

A quick detection method for the early stage of disease in turbot(Scophthalmus maximus )-Development of the colloidal gold-based immunochromatographic strip
Wang Weifang,Chai Shujun,Liu Qingtang,Lei Jilin,Ding Fuhong,Hong Lei,Liu Xinfu and Su Ke.A quick detection method for the early stage of disease in turbot(Scophthalmus maximus )-Development of the colloidal gold-based immunochromatographic strip[J].Engineering Science,2012,14(2):8-13.
Authors:Wang Weifang  Chai Shujun  Liu Qingtang  Lei Jilin  Ding Fuhong  Hong Lei  Liu Xinfu and Su Ke
Affiliation:1.Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China;2.Henan Key Laboratory for Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002, China;3.Qingdao General Aquaculture Co.,Ltd.,Qingdao 266404,China)
Abstract:In this study, an immunochromatographic strip was developed for the serological detection of antibodies against bovine serum albumin (BSA) in turbot (Scophthalmus maximus), in which the BSA was taken as antigen. In the strip, the rabbit anti turbot immunoglobulin M, made from this research, labeled with colloidal gold was used as the detector, and the BSA and staphylococcal protein A were blotted on the nitrocellulose membrane for the test and control lines, respectively. The positive serum samples from turbot vaccinated with BSA and negative serum samples from normal turbot were collected to evaluate the characteristics of the strip, and the results were compared with the method of enzyme linked immunosorbent assay (ELISA) established in this study. The strip was shown to be of high specificity and sensitivity, and similar to ELISA. Furthermore, this dipstick assay based on the strip is rapid (5 min) and easy to perform with no requirement of professional skills, reagents or equipments. This suggests that the immunochromatographic strip is an acceptable alternative for use in clinical laboratories lacking specialized equipment and for field diagnosis. Based on the model of this strip developed in the study, this pattern of strip could be performed into any antibody detection in aquaculture animals; it provides a rapid and simple test method to validate the efficacy of the vaccine and to detect and monitor the fish disease in time.
Keywords:turbot(Scophthalmus maximus)  antibody  immunochromatographic strip  point-of-care test  colloidal gold  disease
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