title: Isothermal amplification detection of miRNA based on the catalysis of nucleases and voltammetric characteristics of silver nanoparticles.
abstract: MiRNAs are a fascinating kind of biomolecule due to their vital functions in gene regulation and potential value as biomarkers for serious diseases including cancers. Exploiting convenient and sensitive methods for miRNA expression assays is imperative. In this study, we employ an exonuclease (RecJf) and a nicking endonuclease (Nt.BbvCI) to catalyse isothermal reactions for the amplified detection of miRNA. The degree of cyclical enzymatic amplification depends on the initial target miRNA level, which can determine the density of DNA probes bound on the electrode surface. Since DNA probes with an amino group at the 3' end are able to locate silver nanoparticles on the electrode, which provide intense stripping responses, the sensitive quantification of miRNA can be achieved. The proposed method has a limit of detection as low as 35 aM, with remarkable specificity, which offers a new approach for investigating miRNA networks and for clinical diagnosis applications.
authors: Department of Laboratory Science, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, P. R. China.
authors: Jianhua
authors: J
authors: Xu
authors: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, P. R. China. miaopeng@sibet.ac.cn.
authors: Kun
authors: K
authors: Han
authors: Department of Laboratory Science, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, P. R. China.
authors: Dongdong
authors: D
authors: Liu
authors: Department of Laboratory Science, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, P. R. China.
authors: Li
authors: L
authors: Lin
authors: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, P. R. China. miaopeng@sibet.ac.cn.
authors: Peng
authors: 0000-0003-3993-4778
authors: P
authors: Miao
mesh_terms: D015374:Biosensing Techniques; D002384:Catalysis; D055664:Electrochemical Techniques; D004566:Electrodes; D004720:Endonucleases / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D053758:Nanoparticles / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques; D012680:Sensitivity and Specificity; D012834:Silver
chemical_list: D035683:MicroRNAs; D012834:Silver; D004720:Endonucleases