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本文建立了一种基于辣根过氧化物酶(HRP)和碱性磷酸酯酶(ALP)化学发光底物分辨的双组分免疫分析新技术,用以检测人胶质瘤血清标志物神经元特异性烯醇化酶(NSE)和糖链抗原15-3(CA15-3).实验详细考察了捕获抗体、检测抗体、HRP和ALP酶标记物的用量,结果发现NSE和CA15-3的线性范围分别为0.5～20ng/mL(R2>0.99)和0.5～20U/mL(R2>0.99),最低检出限分别为0.2ng/mL和0.2U/mL;高、中和低三个浓度的血清加样回收率良好;天内和天间相对标准偏差均小于10%;且一份血清,两组分同时检测无交叉干扰.综合而言:本法能够一次实验,高灵敏、高特异地同时检测两种疾病标志物,具有血清用量少、检测时间短、操作简单、结果可靠的优点,有望为胶质瘤的临床早期诊断提供坚实的支持.
In this paper, a novel two-component immunoassay based on HRP and ALP chemiluminescent substrates was developed to detect human glioma serum marker neurons Specific enolase (NSE), and carbohydrate antigen 15-3 (CA15-3) .Experimental details of capture antibody, detection antibody, HRP and ALP enzyme markers were used and the linear range of NSE and CA15-3 was found (R2> 0.99) and 0.5 ~ 20U / mL (R2> 0.99), the detection limits were 0.2ng / mL and 0.2U / mL, respectively. The serum concentrations of high, The sample recovery rate is good; the relative standard deviations in both days and days are less than 10%; and one serum and two components are tested simultaneously without cross interference. In summary, this method can be tested simultaneously, highly sensitive and highly specific The two disease markers, with less serum, shorter detection time, simple operation and reliable results, are expected to provide solid support for early clinical diagnosis of glioma.