重组汉坦病毒核蛋白抗原用于免疫滴金技术检测肾综合征出血热抗体的研究

目的　探索应用重组汉坦病毒核蛋白 (rNP)的免疫滴金法 (CGIDA)对肾综合征出血热 (hemorrhagicfeverrenalsyndrome ,HFRS)的诊断价值。 方法　制备纯化汉坦病毒核蛋白抗原 ,构建了HTN型汉坦病毒的核心区域 (334个碱基 ) ,克隆至原核表达载体 pGEX 4T 1中进行原核表达及纯化。在相同的CGIDA系统中 ,比较研究rNP与天然汉坦病毒核蛋白 (NP)的抗原功能。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　用rNP和天然NP平行检测HFRSIgM符合率达 89.7% ,HFRSIgG符合率达92 .3% ;CGIDA法检测HFRSIgM的敏感性为 75 .0 % ,特异性为 10 0 % ;CGIDA法检测HFRSIgG的敏感性为 83.1% ,特异性为10 0 %。结论　重组核蛋白的CGIDA对HFRS的早期诊断具有较好的应用价值。

Recombinant nucleocapsid protein of Hantavirus as antigen for colloidal gold immuno-dot assay to detect hemorrhagic fever renal syndrome

Objective To use recombinant nucleocapsid protein (rNP) of Hantavirus as antigen in colloidal gold immuno dot assay(CGIDA) for the detection of IgM and IgG antilbodies in hemorrhagic fever renal syndrome(HFRS). Methods The antigen of rNP was prepared by prokaryotic expression and purification of the core sequence of the small (S) genome segment of Hanta strain HTN. The same system of CGIDA was used in comparing the effect of recombinant protein with that of natural nucleoprotein from the lyses of cultural cell infected by the same virus. The method was also compared to those of specific antibodies by IFA for IgG and ELISA for IgM. Results The coincidence rate of recombinant protein and natural protein was 89.7% for detecting IgM and 92.3% for IgG. 300 sera from patients 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 IgG was 100%, and the sensitivity was 83.1%. Conclusions Recombinant NP is effective and valuable antigen used in CGIDA for diagnosis of HFRS.