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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

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【作者】 余鹏博王敬军张家驹郝志明刘中华胡兴菊唐青许文波

【Author】 YU Peng-bo,WANG Jing-jun,ZHANG Jia-ju,et al.(Virus Laboratory of Shanxi Provincial Center for Disease Control and Prevention,Xi’ an,710054,Shanxi,China)

【机构】 陕西省疾病预防控制中心西安交通大学中国疾病预防控制中心病毒学研究所中国疾病预防控制中心病毒学研究所 陕西西安710054陕西西安710054北京100176

【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。

【Abstract】 [Objective]Establishing IgM capturing ELISA assay with nucleocapside protein of Hantaan virus and its McAb labeled with horseradish peroxidase.[Methods]Preparation and purification of recombining nucleocapside protein(rNP) antigen(Ag) and its McAb labeled with horseradish peroxidase(HRP),with enzyme substrate system(TMBH2O2) play as a tracer we can detect anti-HFRS virus IgM from sera by means of polystyrene plate coated with the sheep anti-human μ chain.Result compared to that detected by immunofluorescence 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 result from two method has no significant difference.[Conclusion]The IgM capturing ELISA reagent involved recombined nucleocapside protein antigen and its McAb labeled with HRP can be used in detecting sera specific IgM antibody to diagnoses of hemorrhagic fever with renal syndrome in early infection stage.

【基金】 陕西省首届医药卫生重点科研基金项目(99zh-001)
  • 【文献出处】 预防医学论坛 ,Preventive Medicine Tribune , 编辑部邮箱 ,2006年01期
  • 【分类号】R446.6
  • 【下载频次】93
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