测定抗-HBs的两种方法在不同温育时间的结果比较

Comparison of anti-HBs detection results with two methods and different incubation time

目的探讨检测乙型肝炎(下称乙肝)病毒表面抗体(抗-HBs)两种方法在不同温育时间的结果差异。方法采用免疫酶法(EIA)一步法、时间分辨荧光(TRF)法检测抗-HBs,OD值在0.056～0.380之间标本温育时间变化而造成的结果值的差异。结果用30minEIA一步法检测抗-HBs,结果37例弱阳性和59例阴性,再用60minEIA一步法抗-HBs检测,结果41例呈弱阳性和55例阴性。用TRF两步法抗-HBs测定出42例弱阳性和54例阴性,发现有6例标本不同于温育30min的EIA一步法抗-HBs检测的结果。结论酶联免疫吸附试验作为一种固相免疫测定,抗原抗体的结合反应在固相上进行,要使液相中的抗原或抗体与固相上的特异抗体或抗原完全结合,必须在一定的温度、时间条件下反应。温育所需时间与温度成反比,尤其在冬天,标本从室温到恒温箱这一过程很容易造成实际测定温育时间不够,也是造成灰区误差的原因之一。

Objective To discuss the anti-HBs detection results with three different incubation time. Methods Enzyme immunoassay （EIA） one-step method and time-resolved fluoroimmunoassay （TRF） were applied to observing the difference of anti-HBs detection results （OD value： 0. 056-0. 380） with three different incubation time. Results By applying 30-minute one-step anti-HBs EIA, 39 weak positive cases and 59 negative cases were detected. By applying 60-minute EIA one-step anti-HBs test, test results showed 41 cases were weak positive and 55 cases were negative. 42 weak positive cases and 54 negative cases were detected by means of time-resolved fluoroimmunoassay two step method, in which six cases＇ anti-HBs detection result was different from that tested with 30-minute one-step anti-HBs EIA method. Conclusion To ELISA, a solid-phase immunoassay, antigen-antibody reacted on solid phase. To make antigen or antibody in liquid-phase fully reacts with specific antigen or antibody on soil-phase, a proper temperature and incubation time are necessary. Incubation temperature is inversely proportional to the incubation time. Especially in winter, the process that samples are moved from room temperature to thermostat, will lead to the losing of real incubation time, which is also one of the reasons for grey area error.