VEGF-TrFIA及其在结直肠癌中的应用

Preparation of a Time-Resolved Fluoroimmunoassay Kit for VEGF Detection and Its Clinical Application in Diagnosis of Colorectal Cancer

目的:研制VEGF时间分辨荧光免疫分析(TrFIA)试剂盒。方法:采用双抗体夹心法建立VEGF-TrFIA试剂盒,并对试剂盒的各项指标进行评价。结果:试剂盒的线性范围为1pg/m l^400pg/m l,灵敏度为0.21pg/m l,准确度高,批内和批间变异系数分别为1.1%~8.4%,2.97%~11.7%。与CA199,CA215,AFP,CEA和白蛋白无交叉反应。收集30例结直肠癌患者和37例健康人血清,用本试剂盒测得结直肠癌患者血清VEGF浓度(357.3±22.1)pg/m l显著高于健康人(71.5±9.1)pg/m l(P<0.0001)。本试剂盒检测结果与商用ELISA试剂盒检测结果相关系数为0.993。结论:试剂盒各项指标达到规定的要求,可用于临床血清VEGF检测。

Objective To prepare a time - resolved fluoroimmunoassay （TrFIA） kit for detecting human VEGF. Methods A sandwich TrFIA kit for VEGF detection was developed in this laboratory using two strains of anti - VEGF monoclonal antibody. Results This VEGF - TrFIA kit was capable of detecting VEGF within the range of 1～400pg/ml with the sensitivity of 0.21pg/ml and without cross - reactivity with CA199, CA215, AFP, CEA, or albumin. The intra - and inter - assay coefficients of variation were 1.1%～8.4% and 2.9%～11.7%, respectively. Serum samples were collected from 30 patients with colorectal cancer and 37 controls. VEGF contents were significantly higher in colorectal cancer patients than those in controls （357.3pg/ml±22.1pg/ml vs 71.5pg/ml±9.1pg/ml; P 〈 0.0001）. The correlation coefficient between the results from this kit and the conventional commercially available ELISA kit for this series of 67 blood samples was 0.993. Conclusion The prepared TrFIA kit for VEGF detection meets the demand of clinical application with good sensitivity, precision, specificity and accuracy.