title: A novel surface plasmon resonance biosensor for enzyme-free and highly sensitive detection of microRNA based on multi component nucleic acid enzyme (MNAzyme)-mediated catalyzed hairpin assembly.
abstract: MicroRNAs (miRNAs) are potentially useful biomarkers for early diagnosis of human diseases. Here, a simple surface plasmon resonance (SPR) biosensor has been developed for highly sensitive detection of miRNA by designing a new enzyme-free and isothermal amplification strategy, named multi component nucleic acid enzyme-mediated mismatched catalyzed hairpin assembly (MNAzyme-CHA). The partial MNAzymes co-recognized the target to form a stable active MNAzyme, which continued to digest multiple hairpin H0 substrates, concomitantly generating a lot of fragments. The H0 fragments could initiate the mismatched CHA cycles, resulting in the generation of massive hairpin H1-H2 complexes. As a result, the H1-H2 complexes and streptavidin were attached to the sensor surface, leading to a significantly amplified SPR signal readout. The established biosensor showed high sensitivity and selectivity with a wide dynamic range from 1 pM to 100 nM. It was also successfully applied to the determination of target miRNA spiked into human total RNA samples. Thus, this developed biosensing strategy presents a simple and stable platform toward sensitive and convenient miRNA detection, and has great potential in assays of many other nucleic acids analytes for biomedical research and early clinical diagnosis.
authors.affiliation: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xinmin
authors.initials: X
authors.lastname: Li
authors.affiliation: The Center for Clinical Molecular Medical detection, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Cheng
authors.affiliation: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Li
authors.affiliation: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Jiangling
authors.initials: J
authors.lastname: Wu
authors.affiliation: Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Ding
authors.affiliation: Department of Chemistry, University of California, Riverside, CA 92521, United States.
authors.forename: Quan
authors.initials: Q
authors.lastname: Cheng
authors.affiliation: 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.forename: Shijia
authors.initials: S
authors.lastname: Ding
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002384:Catalysis; D006801:Humans; D057230:Limit of Detection; D035683:MicroRNAs / Q000737:chemistry / Q000302:isolation & purification*; D009690:Nucleic Acid Conformation; D009696:Nucleic Acids / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry; D020349:Surface Plasmon Resonance
chemical_list: D035683:MicroRNAs; D009696:Nucleic Acids; D019809:Streptavidin
keywords: MNAzyme; MicroRNA; Mismatched catalyzed hairpin assembly; Streptavidin; Surface plasmon resonance