Du Jie, Liu Jinjuan, Li Hongyan
Department of Health Examination Center, Shaanxi Provincial People Hospital, Xi'an, China.
Anal Sci. 2025 Jun 25. doi: 10.1007/s44211-025-00791-z.
Monitoring the levels of various biomarkers, such as nucleic acids and proteins, is crucial for early disease diagnosis, evaluating treatment efficacy, and predicting patient prognosis, thereby aiding clinical decision-making. Herein a robust double-circuit fluorescence assay combining the advantages of EDC and CHA called EDCHA is proposed. The EDCHA includes two cycles, EDC system identifies the target miRNA, realizes first signal amplification and releases an initiator chain to trigger the CHA system. CHA system realizes second signal amplification by opened hairpin structures and different fluorophore groups. In addition, by replacing the waste chain in the EDC system with fluorescent-labeled aptamer, the EDCHA can simultaneously detect both miRNA and protein. The EDCHA was verified with the specific detection of miRNA-21 with a low detection limit (LOD) of 77.18 pM. Furthermore, our method enables the simultaneous detection of two types of biomarkers-nucleic acids and proteins-with a low detection limit (LOD) of 0.94 nM for the AD-related protein AβO. These results demonstrate the enormous potential of EDCHA in diverse biomarkers analysis and cancer diagnosis.
监测各种生物标志物（如核酸和蛋白质）的水平对于疾病的早期诊断、评估治疗效果以及预测患者预后至关重要，从而有助于临床决策。本文提出了一种结合了EDC和CHA优势的强大双循环荧光检测法，称为EDCHA。EDCHA包括两个循环，EDC系统识别目标miRNA，实现第一次信号放大并释放引发链以触发CHA系统。CHA系统通过打开的发夹结构和不同的荧光团实现第二次信号放大。此外，通过用荧光标记的适体替换EDC系统中的废链，EDCHA可以同时检测miRNA和蛋白质。EDCHA通过对miRNA-21的特异性检测得到验证，检测限（LOD）低至77.18 pM。此外，我们的方法能够同时检测两种类型的生物标志物——核酸和蛋白质，对于与阿尔茨海默病相关的蛋白质AβO，检测限（LOD）低至0.94 nM。这些结果证明了EDCHA在多种生物标志物分析和癌症诊断中的巨大潜力。
