摘要
目的 确定定性酶联免疫吸附试验 (ELISA)测定中温育温度、时间和方式对测定结果的影响程度。方法 温育温度分别设为 3 7℃、43℃和室温 ,温育时间分别为 45分钟、60分钟、75分钟、90分钟和 2小时。其中室温温育又分别采用振荡和静置两种条件 ,温育时间分别为 45分钟、1小时、2小时和 4小时。检测阴性血清和 0 .2、0 .5、1、2、5ng/mlHBsAg系列定值血清 ,观察这些条件下标本测定S/CO比值的差异。结果 在同一温育温度下 ,所有血清的S/CO比值均随着测定温育时间的延长而增大 ,浓度越低表现越为明显 ,从 0 .2ng/ml到 5ng/ml的样本 ,温育 2小时的S/CO比值与温育 1小时相比 ,均有明显的增加 ,0 .2ng/ml的样本由按CUT OFF值判断为阴性而转为阳性。而同时阴性血清的S/CO比值则变化不大。在振动状态下反应较之静止状态下的反应 ,S/CO比值均有明显增加。结论 温育条件对ELISA测定结果有很大影响 ,尤其是弱阳性标本。
Objective To determine the effects of incubation temperature , time and mode on results of HBsAg by ELISA. Methods A negative serum sample and a series of sera samples with different HBsAg concentration (0.2, 0.5, 1.0, 2.0, 5.0 ng/ml) were tested by using commercial ELISA kits under conditions set as follows: incubation time was set for 45, 60, 75, 90 and 120 minutes when incubation temperatures were at 37℃,43℃ and room temperature(23±1(C). Two incubation modes ( static and shaking) were used for room temperature incubation.The times for the incubation set as 45min,1hr,2hrs and 4 hrs. Results With incubation time extended, the S/CO values of the sera with HBsAg increased, especially for weak positive samples. Compared two hours and one hour incubation time, there was a significantly increase in S/CO value of the series of the sera with HBsAg. Moreover, the negative results of the sample with 0.2 ng/ml HBsAg when used one hour incubation changed to positive with two hours incubation. Meanwhile, there was no markedly changes in S/CO values of serum without HBsAg. The S/CO values of the series of sera with HBsAg at room temperature with shaking were much higher than those without shaking. Conclusion The results showed that incubation conditions markedly influences S/CO values of ELISA, especially on weak positive samples.
出处
《实用肝脏病杂志》
CAS
2005年第1期16-18,共3页
Journal of Practical Hepatology
