title: A simple and highly sensitive DNAzyme-based assay for nicotinamide adenine dinucleotide by ligase-mediated inhibition of strand displacement amplification.
abstract: Existing strategies for detecting nicotinamide adenine dinucleotide (NAD(+)) or other cofactors are commonly cumbersome and moderate sensitive. We report a novel DNAzyme-based visual assay strategy for NAD(+) based on ligase-mediated inhibition of the strand displacement amplification (SDA). In the presence of NAD(+), the SDA can be inhibited by the ligase reaction of two primers, which can initiate the SDA reaction in the case of no ligation, resulting in a dramatically decreasing yield of the SDA product, a G-quadruplex DNAzyme that can quantitatively catalyze the formation of a colored product. Therefore, the quantitative analysis for NAD(+) can be achieved visually with high sensitivity. The developed strategy provides a simple colorimetric approach with high selectivity against most interferences and a detection limit as low as 50 pM. It also provides a universal platform for investigating cofactors or other related small molecules as well as quantifying the activity of DNA ligases.
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Ying-Ya
authors.initials: YY
authors.lastname: Kan
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: jianhuijiang@hnu.edu.cn.
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: rqyu@hnu.edu.cn.
authors.forename: Ru-Qin
authors.initials: RQ
authors.lastname: Yu
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D021881:DNA, Catalytic / Q000378:metabolism*; D008025:Ligases / Q000378:metabolism*; D057230:Limit of Detection; D009243:NAD / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D021881:DNA, Catalytic; D009243:NAD; D008025:Ligases
keywords: Inhibition; Ligase-mediated; Nicotinamide adenine dinucleotide; Strand displacement amplification