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

To develop a comprehensive detection platform for immunoprotection of inactivated coxsackievirus A16(CA16) vaccine based on the neutralizing antibodies tested through competitive inhibition ELISA, micro-cytopathic effect neutralization test and neonatal mice challenge protection test.

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 taken at 0, 4, 6, 8, 10 weeks and serum neutralizing 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.

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 inhibition-ELISA and neonatal mice challenge protection test and 0.89 between micro-cytopathic effect neutralization test and neonatal mice challenge protection test.

A comprehensive detection system through competitive inhibition ELISA, micro-cytopathic effect neutralization test and neonatal mice challenge protection test measuring the neutralizing antibodies 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.