摘要
目的通过探讨钩状效应对酶联免疫吸附试验(ELISA)双抗原夹心一步法检测梅毒特异性抗体结果的影响,评价双抗原夹心ELISA一步法检测梅毒螺旋体特异性抗体的应用价值。方法采用稀释法,ELISA双抗原夹心二步法和振荡法分别检测怀疑存在钩状效应的血清标本8例;采用ELISA双抗原夹心一步法和二步法分别检测40例健康体检者血清和卫生部临床检验中心提供的梅毒抗体临床科研血清盘一套。结果8例ELISA双抗原夹心一步法检测结果为阴性,怀疑存在钩状效应的患者血清标本,采用ELISA双抗原夹心二步法进行检测时,结果均转为阳性;进行1∶2,1∶10,1∶50和1∶100倍稀释后,采用ELISA双抗原夹心一步法进行检测,分别有7例,8例,8例和5例结果转为阳性;将反应板在孵育过程中同时进行振荡,有5例转为弱阳性;ELISA双抗原夹心一步法和二步法分别检测40例健康体检者血清和卫生部临床检验中心提供的梅毒抗体临床科研血清盘,除3例弱阳性结果不符外,其它结果均一致。结论建立敏感度高的ELISA两步法或采用现有ELISA一步法试剂同时对标本进行原倍和一定倍数稀释后检测,可以既克服钩状效应,又保证检测的敏感度。
Objective to investigate if there was high dose hook effect in measuring anti-syphilis specific antibody by one-step enzyme-linked immunosorbent assay(ELISA),influence and overcome methods.Methods Firstly,anti-syphilis specific antibody in 40 sera from healthy people,40 serum panels from the National Center for Clinical Laboratory and 8 sera from the patients clinically diagnosed as syphilis were measured by one-step ELISA separately.Secondly,After 1:2,1:10,1:50 and 1∶100 diluted,8 sera from patients with syphilis were detected again by onestep ELISA only.Thirdly,8 sera were detected by one-step ELISA again through shaking the microplate in 200 times/min during incubation.Results The results of 4 weak positive samples from the serum panels detected by two-step ELISA were negative;The results of 8 samples from patients with syphilis,no positive result was found by one-step ELISA,were all positive detected by two-step ELISA and 7,8,8,and 5 positive respectively detected by one-step ELISA again after diluted in 1∶2,1∶10,1∶50 and 1∶100.5 of 8 negative samples detected by one-step ELISA showed weak positive when the microplate was mixed by shaking during incubation.Conclusion There was high dose hook effect in measuring anti-syphilis specific antibody by one-step ELISA.The hook effect can be overcome by establishing a new two-step ELISA with high specificity and high sensitivity or diluting all samples.
出处
《中国实验诊断学》
2007年第11期1520-1523,共4页
Chinese Journal of Laboratory Diagnosis
关键词
酶联免疫吸附实验
钩状效应
一步法
梅毒螺旋体抗体
enzyme-linked immunosorbent assay
hook effect
One step ELISA
anti-syphilis specific antibody
