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作者

Xin Jin Jiankang Zhu Yun Zhang Shuaichen Jin Xinxin Zhao Qian Xu Dexu Zhang Linchuan Li Qiongzheng Hu Guangyong Zhang

作者单位

³Laboratory of Metabolism and Gastrointestinal Tumor, The First Affiliated Hospital of Shandong First Medical University, Jinan 250014, China ⁵School of Pharmaceutical Sciences, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250014, China ²Department of General Surgery, The First Affiliated Hospital of Shandong First Medical University, Jinan 250014, China ⁴Qilu University of Technology (Shandong Academy of Sciences), Shandong Analysis and Test Center, Jinan 250014, China ¹Department of General Surgery, Shandong Provincial Qianfoshan Hospital, Cheeloo College of Medicine, Shandong University, Jinan 250014, China

刊名

Microchemical Journal

年份

2025

卷号

Vol.208

页码

112317

ISSN

0026-265X

摘要

Detecting tumor biomarkers in blood samples is essential in cancer diagnosis and prognosis. This study proposes a CRISPR/Cas system-mediated colorimetric assay to detect the tumor biomarker in human serum samples. It involves using the tumor biomarker as a switch of the CRISPR/Cas system to modulate the function of Hg, which is a typical urease inhibitor. The detection of carcinoembryonic antigen is demonstrated as an example. The CEA aptamer is used to activate the CRISPR/Cas system, allowing ...更多

Detecting tumor biomarkers in blood samples is essential in cancer diagnosis and prognosis. This study proposes a CRISPR/Cas system-mediated colorimetric assay to detect the tumor biomarker in human serum samples. It involves using the tumor biomarker as a switch of the CRISPR/Cas system to modulate the function of Hg, which is a typical urease inhibitor. The detection of carcinoembryonic antigen is demonstrated as an example. The CEA aptamer is used to activate the CRISPR/Cas system, allowing for the cleavage of a single-stranded DNA rich in thymine nucleotides, thereby causing the inhibition of the urease activity by Hg. However, in the presence of CEA, the CRISPR/Cas system cannot be activated due to the specific binding of CEA to its aptamer, which leads to the capture of Hg owing to the formation of a DNA hairpin structure with T-Hg-T coordination bonds. Thus, the urease activity is remained. Since the enzymatic hydrolysis of urea by urease increases the pH of the urea solution containing phenol red and changes the solution from yellow to pink, CEA can be detected with the assistance of the colorimetric assay. This method shows high sensitivity and selectivity for CEA with a detection limit of 0.028 ng/mL. In addition, serum samples from patients with normal and abnormal levels are tested to verify the validity of the proposed method for practical applications. Overall, this study provides a simple, fast, and stable colorimetric assay to detect the tumor biomarker in human serum and has great potential for early cancer screening and prognostic determination.收起

文献类型

期刊