摘要
目的:建立一种简便的血清HBsAg检测二步法标记免疫试验。方法:在既往G6ELISA的基础上建立血清HB-sAg检测排溢法ELISA,采用系列稀释实验对方的相对显色强度,敏感性,及其对Hooks效应的拮抗能力进行考核,将其用于对一组临床标本的检测,并与经典二步法及一步法ELISA进行比较。结果:在大致相同的实验条件下排溢法,二步法及一步法ELISA三者实验信号的强度大致相当;其敏感性分别为19.2、12.6及14.5pg/ml。当被检标本HBsAg浓度在37.5μg/ml时一步法ELISA开始显现Hooks效应,至2400μg/ml时测值已接近阈值;而排溢法及经典二步法ELISA两者对Hooks效应则表现出相似的拮抗。对231例临床标本进行检测,112例高滴度阳性标本(经稀释实验证实)中出现中重度Hooks效应者一步法ELISA17例,占15.2%,而排溢法及其二步法ELISA则未见Hooks效应的影响。结论:在保证实验结果的前提下,排溢法ELISA在操作程度简化上与一步法相当,但可完全克服Hooks现象的干扰。
Objective：To establish a simple two-step labeled immune test to detect the serum HBsAg.Methods：The excluded ELISA was established based on the previous G6 ELISA for serum HBsAg detection,and its relative color strength,sensitivity,and the antagonistic capacity to the Hooks effect were assessed using serial dilutions test.The assay was used to detect a group of clinical specimens,and the results were compared with those of the classic two-step and one-step ELISA.Results：The test signal strength of Excluded ELISA was roughly the same of the other two method,two-step and one-step ELISA in similar experimental conditions.The sensitivity was 19.2 pg/ml,12.6 pg/ml and 14.5 pg/ml,respectively.When HBsAg concentration of the samples was up to 37.5 μg/ml,one-step ELISA began to show the Hooks effect,and the measured values were close to the threshold value when HBsAg up to 2 400 μg/ml;But the excluded ELISA and the classical two-step ELISA both were showed similar antagonistic to the Hooks effect.By use of the three methods to detect 231 clinical samples,the results showed that there were 17 samples with moderate to severe Hooks effects when detected by one-step ELISA,acconunting for 15.2% in 112 samples with high-titer positive samples（confirmed by dilution experiments）,whereas there was no Hooks effect detected by the other two methods.Conclusion：In the premise of guaranteed results,the excluded ELISA is equal to the one-step ELISA in the degree of simplification of operation,but it could completely overcome the interference of Hooks effect.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2010年第12期1110-1113,共4页
Chinese Journal of Immunology
基金
科技部“国家2008年科技重大专项课题:艾滋病和病毒性肝炎等重大传染病防治专项”课题(No.2008ZX10205)
