两孔一步法同时检测HBsAg与抗-HBs的ELISA技术

One Step Incubation ELISA for the Detection of Both HBsAg and Anti-HBs by Same Kit Using Monoclonal Antibody Reagents

以一步夹心法检测HBsAg的ELISA技术为基础,结合中和试验原理,提出了两孔一步法同时检测HBsAg与抗-HBs的ELISA技术。对已知RIA法为HBsAg阳性血清108份、抗-HBs阳性且S/N>10的血清395份、HBsAg与抗-HBs均阴性的血清167份,用本法检测,结果符合RIA法的分别为106份(符合率98.14％),385份(97.46％)和158份(94.61％),对乙肝疫苗免疫前的各类健康人血清424份及经乙肝疫苗三针免疫后1—2个月的血清359份,用本法及RIA法平行测试HBsAg与抗-HBs,结果两法总的符合率分别为89.39％(379份)87.30％(282份)。

A new one step incubation ELISA technique using monoclonal antibody reagents was developed, and both HBsAg and anti-HBs could be tested simultaneously by the same ELISA kit. In this system, one well was used for the detection of HBsAg by one step sanwich technique, another well was used for the examination of anti-HBs by one step incubation neutralization technique. The wells for both tests were coated with the same stain of monoclonal anti-HBs, and other stain of monoclonal anti-HBs was used for conjugation with enzyme, the same conjugate was used in both tests. Of 108 HBsAg positive (RIA) sera were tested by this ELISA system, 106 (98.14%) wers found to be positive, of 395 anti-HBs positive (RIA) sera, 385 (97.46%) were also confirmed to be positive by this ELISA. 167 both HBsAg and anti-HBs negative serum samples (RIA) were determined by this ELISA, results of 158 (94.61%) samples were in agreement with each other. 359 sera from vaccines one month after 'HB vaccination were tested for anti-HBs by both RIA and this ELISA, 323 were found to be positive by RIA but 289 by ELISA. Four hundred and twenty-four healthy individuals were examined for HBsAg and anti-HBs by both RIA and FLISA, the results were highly comparative. The authors consider this ELISA system to be a practical, convenient, specific and sensitive technique.