title: Chemiluminescence imaging for microRNA detection based on cascade exponential isothermal amplification machinery.
abstract: A novel G-quadruplex DNAzyme-driven chemiluminescence (CL) imaging method was developed for ultrasensitive and specific detection of miRNA based on the cascade exponential isothermal amplification reaction (EXPAR) machinery. A structurally tailored hairpin probe switch was designed to selectively recognise miRNA and form hybridisation products to trigger polymerase and nicking enzyme machinery, resulting in the generation of product I, which was complementary to a region of the functional linear template. Then, the response of the functional linear template to the generated product I further activated the exponential isothermal amplification machinery, leading to synthesis of numerous horseradish peroxidase mimicking DNAzyme units for CL signal transduction. The amplification paradigm generated a linear response from 10 fM to 100 pM, with a low detection limit of 2.91 fM, and enabled discrimination of target miRNA from a single-base mismatched target. The developed biosensing platform demonstrated the advantages of isothermal, homogeneous, visual detection for miRNA assays, offering a promising tool for clinical diagnosis.
authors: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors: Yongjie
authors: Y
authors: Xu
authors: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors: Dandan
authors: D
authors: Li
authors: The Center for Clinical Molecular Medical Detection, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors: Wei
authors: W
authors: Cheng
authors: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors: Rong
authors: R
authors: Hu
authors: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors: Ye
authors: Y
authors: Sang
authors: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors: Yibing
authors: Y
authors: Yin
authors: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. Electronic address: dingshijia@163.com.
authors: Shijia
authors: S
authors: Ding
authors: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China; State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210023, China. Electronic address: hxju@nju.edu.cn.
authors: Huangxian
authors: H
authors: Ju
mesh_terms: D015374:Biosensing Techniques; D021881:DNA, Catalytic / Q000378:metabolism; D054856:G-Quadruplexes; D006735:Horseradish Peroxidase / Q000378:metabolism; D008163:Luminescent Measurements; D035683:MicroRNAs / Q000032:analysis* / Q000096:biosynthesis; D021141:Nucleic Acid Amplification Techniques
chemical_list: D021881:DNA, Catalytic; D035683:MicroRNAs; D006735:Horseradish Peroxidase
keywords: Chemiluminescence imaging; Exponential isothermal amplification reaction; Horseradish peroxidase-mimicking DNAzyme; MicroRNA