本研究旨在制备和鉴定小鼠抗Ⅱ型登革病毒(DENV-2)10种蛋白的抗体，为后续相关研究提供实验材料。利用真核表达载体pReceiver构建DENV-2 10种蛋白的重组质粒，提取质粒DNA，肌内注射免疫小鼠，共免疫4次。末次免疫后2周取小鼠血清，利用DENV-2感染的Vero细胞和DENV-2各蛋白的稳定表达细胞，通过酶联免疫吸附试验(ELISA)、间接免疫荧光法(IFA)和蛋白免疫印迹法评价免疫效果，分析抗体的特点。DNA免疫小鼠后获得抗DENV-2 10种蛋白的抗血清，抗体效价波动于1:400～1:16 127之间，以抗E蛋白抗体效价最高，达1:16 127，而抗NS3、NS4b、NS5蛋白抗体效价较低，仅为1:400。利用DENV-2感染的Vero细胞和稳定表达病毒蛋白的EAhy926细胞进行IFA染色，抗DENV-2各蛋白的抗血清均可特异性识别DENV-2抗原。蛋白免疫印迹结果显示，抗E、NS1、NS4b和NS5蛋白抗体能识别热变性蛋白，其他抗血清未呈现阳性反应条带。本研究提示，DNA免疫小鼠所获得的抗DENV-2各蛋白抗体能特异性识别自然感染或模拟自然感染状态下的DENV-2蛋白，可为后续相关研究提供工具，也表明DNA免疫法可作为抗体制备的一种策略。

Eukaryotic expression vector containing different proteins (C, M, E and NS1-­5) of dengue virus serotype 2 (DENV­-2) were constructed and intramuscularly injected directly to mice for developing antibodies against DENV-­2. Two weeks after the fourth immunization, sera were collected and the immune effects were evaluated by enzyme­linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA) and Western blotting. Specific antibodies could be obtained by DNA immunization, and the titers of these antibodies fluctuated between 1:400­ - 1:16 127. Of those, the antibody against protein E showed the highest titer at 1:16 127 whereas the antibodies against proteins NS3, NS4b and NS5 had the lowest titer at 1:400. The results showed that all antibodies acquired by DNA immunization could produce specific immunofluorescent reactions when tested using DENV­-2-infected Vero cells, suggesting that antibodies prepared by DNA immunization could specifically identify naturally infected DENV­-2. Antibodies against proteins E, NS1, NS4b and NS5 could also identify heat­denatured proteins. The results generated from this study provided the base for further research.