Simultaneous Homogeneous Fluorescence Detection of AFP and GPC3 in Hepatocellular Carcinoma Clinical Samples Assisted by Enzyme-Free Catalytic Hairpin Assembly
Simultaneous Homogeneous Fluorescence Detection of AFP and GPC3 in Hepatocellular Carcinoma Clinical Samples Assisted by Enzyme-Free Catalytic Hairpin Assembly
复制标题
无酶催化发夹组装辅助同时均相荧光检测肝细胞癌临床样品中的 AFP 和 GPC3
DOI：
10.1021/acsami.2c09135
复制
发表时间：
2022-06-14
影响因子：
9.5
通讯作者：
Chen, Jie
中科院分区：
文献类型：
--
作者：
Chen, Piaopiao;Jiang, Pengjun;Chen, Jie
同时检测多个肿瘤标志物具有高灵敏度和高性价比，在癌症诊断中显示出巨大的潜力。在这里，我们报道了一种简单的无酶平行催化发夹组件(CHA)扩增策略，以N-甲基中卟啉IX(NMM)和量子点(Qds)为信号报告分子，用于甲胎蛋白(AFP)和GPC3(GPC3)的均相荧光同时检测。在选择性结合时，两适配子双链DNA(DsDNA)探针释放的单链DNA(SsDNA)触发CHA扩增，进一步释放G-四链序列和C-Ag+-C结构中的Ag+。然后，NMM和CdTe量子点分别选择性地识别G-四链体和Ag+。在优化条件下，AFP和GPC3的检出限分别为3fg/m L和0.25fg/m L。使用基于颜色和距离的视觉读数，GPC3的LOD为1fg/mL。应用该方法对41例肝细胞癌患者的临床血清标本进行了AFP和GPC3的定量检测。AFP和GPC3的定量检测结果与电化学发光免疫分析(ECL-IA)临床试剂盒检测结果一致，并与放射学和病理结果相一致。临床试验结果证实了GPC3作为肿瘤生物标志物的潜力，我们建议肝细胞癌的临界值为2 ng/mLGPC3。
Simultaneous sensitive and cost-effective detection of multiple tumor markers has shown great potential for cancer diagnostics. Herein, we reported a simple enzyme-free parallel catalytic hairpin assembly (CHA) amplification strategy with N-methyl mesoporphyrin IX (NMM) and quantum dots (QDs) as signal reporters for the homogeneous fluorescent simultaneous detection of alpha-fetoprotein (AFP) and glypican-3 (GPC3). Upon selective binding, the released single-stranded DNA (ssDNA) from the two-aptamer double-stranded DNA (dsDNA) probes triggers CHA amplification, further releasing the G-quadruplex sequence and Ag+ from the C-Ag+-C structures at the same time. Then, NMM and CdTe QDs selectively recognize G-quadruplex and Ag+, respectively. Under optimized conditions, limits of detections (LODs) as low as 3 fg/mL for AFP and 0.25 fg/mL for GPC3 were achieved using fluorescence readout. Using color- and distance-based visual readouts, an LOD of 1 fg/mL for GPC3 was reached. This method was applied to quantitatively analyze AFP and GPC3 in 41 clinical serum samples of hepatocellular carcinoma (HCC) patients. The quantitative test results for AFP and GPC3 were consistent with those obtained using the electrochemiluminescence immunoassay (ECL-IA) clinical kit and correlated with radiological and pathological findings. The results of clinical tests demonstrated the potential of GPC3 as a tumor biomarker, and we propose a cut-off value of 2 ng/mL GPC3 for HCC.