【摘要】 目的:基于滚环扩增技术(rolling circle amplification,RCA)结合免疫反应建立一种蛋白质的高灵敏检测方法。方法:根据双抗体夹心法原理,以链酶亲和素为桥梁,生物素修饰的单链寡核苷酸引物可以高亲和力地链接到生物素标记的免疫复合物上,建立蛋白质的滚环扩增技术检测平台,并将其用于血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)的检测。结果:在优化实验条件下,对VEGF检测的灵敏度达到10fM。结论:与常规的酶标二抗体系ELISA法相比,本方法具有更高的灵敏度,适于低丰度蛋白质的检测,此方法的进一步完善将为疾病新的蛋白标志物的筛选以及临床诊断提供有力的工具。更多还原

【Abstract】 Objective: Combined with rolling circle amplification(RCA) and immune response to set up a high throughput test methods of protein.Method: According to the streptavidin as a bridge,the multiplex anchoring of biotinylated single-strand oligonucleotide will bound to the biotinylated immunocomplex.Then set up a new detection method base on the RCA and use the vascular endothelial growth factor(VEGF) as a model protein to verify the practicability of the proposed strategy.Result: In the optimized experimental conditions,the RCA strategy show a limit of detection of about 10 fM.Conclusion: Compare to the conventional ELISA,this method show a higher sensitivity and could meet low abundance protein detection.This strategy would become a powerful tool for screening new disease biomarkers and clinical diagnostics.更多还原