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目的比较不同免疫方案制备肠道病毒71型(EV71)抗血清的免疫效果,选择最佳免疫方案。方法制备EV71,并对病毒进行滴定和灭活。将BALB/c鼠随机分为5组,每组9只。1、2组以灭活病毒免疫,3、4组以活病毒免疫,1、3组的免疫间隔时间为2周,2、4组为3周,第5组为对照组,用磷酸盐缓冲液(PBS)免疫,免疫间隔2周。免疫4次后用间接酶联免疫吸附试验(ELISA)法检测各组小鼠血清抗体滴度。结果 4个实验组均产生阳性抗体(A/A0≥2.1),对照组抗体阴性(A/A0<2.1),1～5组吸光度值依次为1.081 7±0.289 7、1.029 6±0.3719、1.074 8±0.374 9、1.492 9±0.215 9、0.168 7±0.021 8。对其进行单因素方差分析,F=23.449,P<0.05,差异有统计学意义,LSD-t检验结果,第4组与1、2、3组差异均有统计学意义(P<0.05),而1、2、3组两两比较差异无统计学意义(P>0.05)。结论各组抗体滴度不完全相同,其中第4组即活病毒间隔3周免疫组抗体滴度最高,所以活病毒间隔3周免疫方案可作为制得高滴度EV71抗血清的理想选择。
Objective To compare the immunogenicity of antisera against enterovirus 71 (EV71) prepared by different immunization protocols and select the best immunization protocol. Methods EV71 was prepared and the virus titrated and inactivated. BALB / c mice were randomly divided into 5 groups, 9 in each group. Groups 1 and 2 were immunized with inactivated virus, groups 3 and 4 were immunized with live virus, immunization interval of groups 1 and 2 was 2 weeks, groups 2 and 4 were 3 weeks, group 5 was control group, Liquid (PBS) immunization, immunization interval of 2 weeks. After four times of immunization, the serum antibody titers of each group were detected by indirect enzyme-linked immunosorbent assay (ELISA). Results All the four experimental groups had positive antibody (A / A0≥2.1) and negative control group (A / A0 <2.1). The absorbance values of 1-5 groups were 1.081 7 ± 0.289 7, 1.029 6 ± 0.3719 and 1.074 8 ± 0.374 9,1.492 9 ± 0.215 9,0.168 7 ± 0.021 8. One-way analysis of variance (ANOVA), F = 23.449, P <0.05, the difference was statistically significant. There was significant difference between LSD-t test in group 4 and group 1,2,3 (P <0.05) There was no significant difference between groups 1,2,3 (P> 0.05). Conclusion The titer of antibody in each group is not exactly the same. The antibody titer in the fourth group is the highest in the immunized group at the interval of 3 weeks. Therefore, the immunization program with 3 weeks interval of live virus can be used as the ideal choice for preparing the high titer of EV71 antiserum.