结合三螺旋分子探针和催化发夹组装反应对甲胎蛋白的快速无酶比色分析  

Rapid enzyme-free colorimetric analysis of alpha-fetoprotein by combining triple-helix molecular probe and catalytic hairpin assembly reaction

作　　者：翁琴 董严 白刚 雷小东 余华 WENG Qin;DONG Yan;BAI Gang;LEI Xiaodong;YU Hua(School of Medicine and Life Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu,Sichuan 611137,China;Department of Oncology,First Affiliated Hospital of Army Military Medical University,Chongqing 400038,China;Department of General Surgery,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu,Sichuan 610072,China)

机构地区：[1]成都中医药大学医学与生命科学学院,成都611137 [2]陆军军医大学第一附属医院肿瘤科,重庆400038 [3]成都中医药大学附属医院普外科,成都610072

出　　处：《重庆医学》2024年第19期2892-2897,2906,共7页Chongqing Medical Journal

摘　　要：目的构建一种基于三螺旋分子探针(THMP)和催化发夹组装(CHA)反应的新型无酶比色生物传感器(T-CHA),用于低丰度甲胎蛋白(AFP)的可视化检测和精准定量分析。方法通过凝胶电泳以及紫外-可见分光光度计记录AFP诱导T-CHA体系产生的吸光度,以评估T-CHA策略的可行性。分别对THMP和CHA的摩尔比、CHA的反应时间、温度及缓冲液pH值等参数进行了优化。用不同浓度AFP诱导T-CHA发生显色反应,以探究T-CHA的检测效能。结果在最佳实验条件下,不同浓度AFP的对数值在5 pg/mL至10 ng/mL呈现线性关系,检测限为2.29 pg/mL。T-CHA体系在105 min内即可完成对低丰度AFP的精准定量分析及无标记的可视化检测,且T-CHA的灵敏度优于ELISA。结论利用THMP的低背景泄漏和CHA的高催化效率,T-CHA体系可以实现肝细胞癌的早期精准诊断,且在即时检验中具有一定的应用前景。Objective To construct a novel enzyme-free colorimetric biosensor(T-CHA)based on triple-helix molecular probe(THMP)and catalytic hairpin assembly(CHA)reaction for visual detection and precise quantitative analysis of low-abundance alpha-fetoprotein(AFP).Methods The absorbance produced by AFP induced T-CHA system was recorded by gel electrophoresis and ultraviolet-visible spectrophotometer to evaluate the feasibility of T-CHA strategy.The molar concentration ratio of THMP and C HA,reaction time of CHA,temperature and pH value of buffer solution were optimized respectively.The color reaction of T-CHA was induced by different concentrati ons of AFP to explore the detection efficiency of T-CHA.Results Under the optimal experimental conditions,the logarithmic values of AFP at different concentrations showed a linear relationship from 5 pg/mL to 10 ng/mL,and the detection limit was 2.29 pg/mL.The T-CHA system could complete accurate quantitative analysis of low abundance AFP and visual free-labeled detection within 105 min,moreover the sensitivity of T-CHA was superior to that of ELISA.Conclusion Utilizing the low background leakage of THMP and the high catalytic efficiency of CHA,the T-CHA system could realize the early accurate diagnosis of hepatocellular carcinoma,moreover which has a certain application prospect in the real time detection.

关 键 词：肝细胞癌 甲胎蛋白 三螺旋分子探针 催化发夹组装 比色分析 

分 类 号：R735.7[医药卫生—肿瘤]