Two-stage nicking enzyme signal amplification combined with DNAzyme amplification for the detection of bone morphogenetic protein 6 mRNA.
Highly sensitive detection of bone morphogenetic protein 6 (BMP6) mRNA is essential to monitor bone regeneration in the regenerating defects. In this work, we proposed a quantitative approach based on two-stage nicking enzyme signal amplification (NESA) and DNAzyme amplification for highly sensitive detection of BMP6 mRNA. The two-stage NESA involves two templates and two-stage amplification reactions under isothermal conditions. The first template contains two repeat sequences that could hybridize to the target RNA, triggering an exponential amplification. The amplified product was a short single-stranded DNA with the same sequence as the target RNA. The single-stranded DNA can trigger another linear NESA and produces a large amount of horseradish peroxidase (HRP)-mimicking G-quadruplex DNAzyme. This proposed assay showed a quantitative analysis of BMP6 mRNA in a wide range from 1 fM to 100 nM with a detection limit of 0.01 fM.
Department of Orthopaedics, Liuzhou People's Hospital.
Zhaohui
Z
Hu
Yanhong
Y
Zhou
Xiangtao
X
Xie
Renjie
R
Jiang
Ningning
N
Li
D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D055418:Bone Morphogenetic Protein 6 / Q000235:genetics*; D021881:DNA, Catalytic / Q000235:genetics* / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism
D055418:Bone Morphogenetic Protein 6; D021881:DNA, Catalytic; D012333:RNA, Messenger; D003850:Deoxyribonuclease I