重组核衣壳蛋白免疫滴金检测肾综合征出血热抗体差异的研究  

The clinical verification of Colloidal Gold Immuno-Dotting with Recombined Nucleocapsid Proteins detecting HFRS antibody

作　　者：储峰 裴彬 李仲平 周欣[2] 王霞明 

机构地区：[1]上海市南汇区南华医院,上海201300 [2]上海市疾病预防控制中心,上海200336

出　　处：《世界感染杂志》2008年第3期191-194,198,共5页World Journal of Infection

摘　　要：目的比较重组核蛋白抗原制备的胶体金免疫试剂盒与病毒悬液抗原制备的胶体金免疫试剂盒在检测肾综合征出血热特异性抗体的差异。方法以汉坦76—118病毒M14626株s片段为模板，克隆334bp片段，重组表达核衣壳蛋白，以此蛋白作为抗原，制备胶体金试剂盒，与以病毒悬液为抗原的胶体金试剂盒进行平行检测300例肾综合征出血热血清IgM和IgG抗体。并与酶联免疫吸附试验（ELISA）和间接免疫荧光法（IFA）对比检测。结果成功构建了重组核蛋白原核表达载体pGEX-4T-1-S，获得高纯度的抗原，分子量约12kD，以此蛋白为抗原制备胶体金试剂盒。检测300例HFRS病人血清抗HFRSigM，rNPCGIDA阳性检出率为90．7％，后者高于前者，差异有显著性（X^2=32．9，P〈0．05）。用rNP与天然NP平行检测蛋白表达法HFRSIgM76.3％，HFRSIgG84．2％；细胞培养法HFRSIgM86．8％，HFRSIgG92．1％。HFRSIgM符合率达89．7％，HFRSIgG符合率达92．3％；CGIDA法检测HFRS IgM的敏感性为75.0％，特异性为100％；CGIDA法检测HFRSIgG的敏感性为83．1％，特异性为100％。结论该重组核蛋白的CGIDA对HFRS的早期诊断具有较好的应用价值。但是敏感性及特异性均低于细胞培养法的CGIDA。Comparing Colloidal Gold Immuno-Dotting Kit precipitated by recombination NP and inartificial NP in detecting HFRS antibody. In order to construct a method used in early diagnosis and supervision and especially adapt to the basic units, we make use of recombinant protein of NP in CGIDA and establish the detection method of CGIDA. We prepared CGIDA detection reagent, and establish the same system of CGIDA compared with the HFRS acute patiens＇ blood by using of recombination NP and inartificial NP. The method was also compared th those of specific antibodies by IFA for IgG and ELISA of IgM. The results shows： The coincidence rate of recombinant protein and natural protein was 89.7% for detecting IgM and 92.3% for IgG 300 sera from pateints with HFRS and 100 sera from patients of other diseases were tested by CGIDA, IFA and ELISA. When compared with ELISA, the specificity of CGIDA for detection of IgM was 100%, and the sensitivity was 75.0%. Compared with IFA, the specificity of CGIDA for detection of IgM was 100%, and the sensitivity was 83.1%. So we get the conclusion that recombiant NP is effective and valuable antigen used in CGIDA for diagnosis of HFRS.

关 键 词：肾综合征出血热 核衣壳蛋白 基因重组 免疫滴金 检测 

分 类 号：R392.7[医药卫生—免疫学] R512.8[医药卫生—基础医学]