论文部分内容阅读
目的：利用特异性与灵敏性较高的神经元特异烯醇化酶（ NSE）、细胞角蛋白21?1片段（ CYFRA21?1）肺癌相关标志物，研制肺癌双显色的体外联合诊断酶联免疫反应（ ELISA）试剂盒。方法将NSE、CYFRA21?1两种抗体按一定比例组合包被到酶标板的板底制成试剂盒。分别以NSE、CYFRA21?1抗原为研究对象，对包被液、包被时间、包被温度因素进行考察，以试剂盒的稳定性、灵敏度、准确性为指标进行结果分析。结果 NSE、CYFRA21?1肺癌双显色ELISA诊断试剂盒4℃放置时有效期为6个月。试剂盒灵敏度为83?8％（129／154），特异度为53?8％（57／106），准确性为71?5％（186／260），受试者工作特征曲线（ ROC）曲线下面积Az 为0?90，各项指标均较单独的NSE、CYFRA21?1明显提高。结论成功制备双显色ELISA肺癌诊断试剂盒，该试剂盒可提高肺癌检查的准确性、灵敏度，降低假阳性率，具有开发为新型肺癌诊断试剂盒的条件。“,”Objective To develop and verify a diagnostic enzyme?linked immunosozbent assay( ELISA) kit in vitro for lung cancer by using lung cancer?related markers of neuron?specific enolase(NSE) and cytokeratin 19 fragment(CYFRA21?1) with high specificity and sensitivity. Methods In vitro diagnostic ELISA kits were prepared by coating anti?NSE and anti?CYFRA21?1 antibodies at the bottom of the microtiter plates at a proper proportion. NSE and CYFRA21?1 antigens were used as research subjects. Coating buff?er, packet time and packet temperature were investigated by detecting the index of stability, sensitivity and accuracy of ELISA kits. Re?sults Period of validity of in vitro NSE and CYFRA21?1?based dual?color diagnostic ELISA kit for lung cancer was 6 months at 4℃. The sensitivity,specificity and accuracy of in vitro NSE and CYFRA21?1?based, dual?color diagnostic ELISA kit were 83?8%( 129/154),53?8%(57/106) and 71?5%(186/260),which were higher than that of anti?NSE or anti?CYFRA21?1 antibodies diagnostic ELISA kits. And the areas under the receiver operating characteristics curves was 0?90. Conclusion NSE and CYFRA21?1?based lung cancer dual?color diagnostic ELISA kit was successfully prepared, which could improve the sensitivity,specificity and accuracy of de?tecting lung cancer. And it was confirmed that it has the potential of developing into a new diagnostic molecular imaging reagent for lung cancer.