柯萨奇病毒A组16型疫苗免疫后中和抗体检测方法的比较

Comparison of three detection methods for neutralizing antibodies induced by Coxsackievirus A16 vaccine

目的采用3种方法检测灭活的柯萨奇病毒A组16型(CA16)疫苗的免疫保护性抗体水平,旨在构建评价疫苗免疫保护的检测平台。方法 CA16候选株疫苗(3726,4430和4432)按0、4、6、8周,经腹腔接种小鼠、大鼠和豚鼠,采集0、4、6、8周和10周血样。通过竞争抑制ELISA、细胞微孔病变实验和乳鼠攻毒保护实验评估免疫血清中和抗体水平,利用SPSS16.0软件分析3种检测方法相关性。结果血清中和抗体在首次加强免疫2周后达到峰值;竞争抑制ELISA和细胞微孔病变实验测得中和效价的相关系数为0.861;竞争抑制ELISA和乳鼠攻毒保护的相关系数为0.8;细胞微孔病变实验和乳鼠攻毒保护的相关系数为0.89。结论竞争抑制ELISA检测中和抗体与"金标准"相比具有灵敏、快速、大量、低廉等特点,具有广阔的应用前景。

Objective To develop a comprehensive detection platform for immunoprotection of inactivated coxsackievirus A16（CAI6） vaccine based on the neutralizing antibodies tested through competitive inhibition ELISA, micro-cytopathic effect neutralization test and neonatal mice challenge protection test. Methods The female BALB/c mice, SD rats and Hartley guinea pigs were inoculated intraperitoneally at 0, 4, 6, 8 weeks with the three vaccine candidates which were made from 3726, 4430 and 4432 virus absorbed on aluminium adjuvant. Immune sera were iaken at 0, 4, 6, 8, l0 weeks and serum neu- tralizing antibodies were evaluated by competitive inhibition-ELISA, micro-cytopathic effect neutralization test and neonatal mice challenge protection test, respectively. The relationships of the three methods were analyzed by SPSS16.0 statistical software. Results The level of neutralizing antibodies reached the peak after the second booster. The correlation coefficient was 0.861 between competitive inhibition-ELISA and micro-cytopathic effect neutralization test, 0.8 between competitive inhi- bition-ELISA and neonatal mice challenge protection test and 0.89 between micro-cytopathic effect neutralization test and neonatal mice challenge protection test. Conclusion A comprehensive detection system through competitive inhibition ELISA, micro-cytopathic effect neutralization test and neonatal mice challenge protection test measuring the neutralizing anti- bodies in immune sera has been constructed. The competitive inhibition-ELISA＇is superior to ＂golden standards＂ in speed, sensitivity, throughput and price, which may have great application prospect.