摘要
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.
A novel chemiluminescence（CL） substrate-resolved immunoassay platform for the determination of two proteins（NSE and CA15-3） is proposed,based on alkaline phosphatase（ALP） and horseradish peroxidase（HRP） for subsequent CL detection.Several immunoreaction parameters were investigated systematically,including the amount of capture antibody,detection antibody and the dilutions of HRP-SA and anti-FITC-ALP.The dose-response curves showed the linear ranges from 0.5 to 20 ng/mL（R20.99） for NSE and 0.5 to 20 U/mL（R20.99） for CA15-3 with the limits of detection 0.2 ng/mL and 0.2 U/mL,respectively.Serum samples spiked with different amounts of antigen（high、middle、low） were assayed and showed an acceptable spiking recovery.Both the inter-assay and intra-assay variation were less than 10%.Two proteins were determined in a single run and no cross-reaction between assays was found.Overall,this proposed method has been successfully used for the simultaneous determination of dual biomarkers in a single vessel,characteristic of low sample consumption,simple operation with improved assay results and reduced overall cost per assay.This simple and homogeneous technique will find applications in the early diagnosis of human golima.
出处
《中国科学：化学》
CAS
CSCD
北大核心
2010年第6期688-693,共6页
SCIENTIA SINICA Chimica
基金
国家重大基础研究计划(2007CB935800)
教育部新教师基金(200802461096)资助
