柯萨奇病毒B3的多克隆抗体制备  

Preparation of polyclonal antibody of coxsackievirus B3

作　　者：代海丽[1] 栾颖[2] 高铁磊[1] 朱琳[1] 靳占峰[1] 

机构地区：[1]哈尔滨医科大学病理教研室和法医教研室 [2]哈尔滨医科大学附属第二医院心内科,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2011年第5期397-400,共4页Journal of Harbin Medical University

基　　金：黑龙江省自然科学基金项目(D2007-54);黑龙江省教育厅科学技术研究项目(11531079)

摘　　要：目的制备柯萨奇病毒B3(coxsackievirus B3,CVB3)的多克隆抗体。方法针对CVB3基因的VP1区设计特异性引物,RT-PCR扩增VP1基因并将其克隆到pGEX-6p-1中,经IPTG诱导表达后,以GST-VP1融合蛋白作为抗原免疫新西兰大白兔,制备多克隆抗体。采用ELISA、Western blot以及间接免疫荧光检测多克隆抗体的效价和特异性。结果 ELISA检测血清的效价不低于1∶64 000,Western blot和间接免疫荧光显示该抗体具有良好的特异性。结论成功制备了效价较高的CVB3多克隆抗体,可应用于CVB3蛋白的检测。Objective To prepare polyclonal antibody against coxsackievirus B3（CVB3）.Methods Specific primers which based on VP1 gene of CVB3 were designed,VP1 gene amplified by RT-PCR was cloned into pGEX-6p-1.High titer polyclonal antiserum was obtained by immunizing New Zealand rabbits with GST-VP1 fusion protein which was induced by IPTG.Then titer of the polyclonal antibody was determined by ELISA and specificity of it was tested by Western blot and indirect immunofluorescence assay.Results The titer of antiserum against CVB3 was above 1∶ 64 000,Western blot and indirect immunofluorescence analysis showed that the polyclonal antibody could bind to the CVB3 protein specifically.Conclusion High specific polyclonal antibody against CVB3 was prepared,which might be used for detection of CVB3 protein in commercial products.

关 键 词：柯萨奇病毒B3 多克隆抗体 原核表达 融合蛋白 

分 类 号：R373.23[医药卫生—病原生物学]