机构:[a]Department of Laboratory Science, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, P. R. China广东省中医院[b]Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, P. R. China.
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 (RecJ(f)) 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 30 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.
基金:
This work was supported by the National Natural Science
Foundation of China (grant no. 31400847), the Science and
Technology Program of Guangdong (grant no. 2014A020212274)
and the Science and Technology Program of Guangzhou (grant
no. 201510010044).
第一作者机构:[a]Department of Laboratory Science, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, P. R. China
通讯作者:
推荐引用方式(GB/T 7714):
Jianhua Xu,Kun Han,Dongdong Liu,et al.Isothermal amplification detection of miRNA based on the catalysis of nucleases and voltammetric characteristics of silver nanoparticles[J].MOLECULAR BIOSYSTEMS.2016,12(12):3550-3555.doi:10.1039/c6mb00659k.
APA:
Jianhua Xu,Kun Han,Dongdong Liu,Li Lin&Peng Miao.(2016).Isothermal amplification detection of miRNA based on the catalysis of nucleases and voltammetric characteristics of silver nanoparticles.MOLECULAR BIOSYSTEMS,12,(12)
MLA:
Jianhua Xu,et al."Isothermal amplification detection of miRNA based on the catalysis of nucleases and voltammetric characteristics of silver nanoparticles".MOLECULAR BIOSYSTEMS 12..12(2016):3550-3555