汉坦病毒核蛋白及其单抗标记物在建立ELISA试剂中的应用  

Application of Nucleocapside Protein of Hantaan Virus and its McAb Labeled with Horseradish Peroxidase in Preparing of IgM Capturing ELISA Assay

作　　者：余鹏博[1] 王敬军[1] 张家驹[1] 郝志明[2] 刘中华[2] 胡兴菊[1] 唐青[3] 许文波[3] 

机构地区：[1]陕西省疾病预防控制中心,陕西西安710054 [2]西安交通大学,陕西西安710054 [3]中国疾病预防控制中心病毒学研究所,北京100176

出　　处：《预防医学论坛》2006年第1期1-3,共3页Preventive Medicine Tribune

基　　金：陕西省首届医药卫生重点科研基金项目(99zh-001)

摘　　要：[目的]利用重组汉坦病毒核蛋白及其单克隆抗体的酶标记物建立IgM捕获ELISA诊断试剂。[方法]制备、纯化重组核蛋白抗原和抗核蛋白单抗并进行辣根过氧化物酶标记,在抗人μ链包被板中结合四甲基联苯胺-过氧化氢(TMB/H2O2)酶底物系统,从血清中检测肾综合征出血热早期特异性IgM抗体,并与免疫荧光抗体法(IFA)进行比较。[结论]检测42份可疑HFRS病人血清,ELISA法和IFA法均阳性的20份,均阴性的19份;ELISA法阴性、IFA法阳性的1份,ELISA法阳性I、FA法阴性的2份,2种方法的差异无统计学意义(配对χ2=0.08,P>0.05)。[结论]用重组汉坦病毒核蛋白及其单克隆抗体的酶标记物建立IgM捕获ELISA诊断试剂可用于HFRS的早期IgM抗体检测。[Objective]Establishing IgM capturing ELISA assay with nueleocapside protein of Hantaan virus and its McAb labeled with horseradish peroxidase. [Methods]Preparatlon and purification of recombining nucleocapside protein （rNP） antigen （Ag） and its MeAb labeled with horseradish peroxidase （HRP）, with enzyme st, bstrale system（TMB/ H2O2 ） play as a tracer we can detect anti-HFRS virus IgM from sera by means of polystyrene plate coated with the sheep antl-human μ chain. Result compared to that detected by immunofluorescenee antibody assay（IFA）. [Results]Detected 42 positive sera and 42 negative sera samples identified by IFA with this IgM capturing ELISA reagent, it shows that the re suit from two method has no significant difference. [Conclusion]The IgM capturing ELISA reagent involved recombined nueleoeapside protein antigen and its McAb labeled with HRP can be used in detecting sera specific IgM antibody to diagnoses of hemorrhagic lever with renal syndrome in early infection stage.

关 键 词：汉坦病毒 核蛋白 酶标记单抗 IgM捕获ELISA HFRS 

分 类 号：R512.8[医药卫生—内科学]