A cyclic enzymatic amplification method for sensitive and selective detection of nucleic acids.
Based on Exonuclease III (Exo III) and displacing probes, we have developed a Cyclic Enzymatic Amplification Method (CEAM) for sensitive and selective detection of nucleic acids. In this design, the displacing probe is non-fluorescent on its own and cannot be digested by Exo III until displacement hybridization by a target sequence, leading to release of free non-quenched fluorophore. Because a single target sequence can lead to the release and digestion of numerous fluorophore strands from the displacing probe, a remarkable signal amplification is achieved. With this method, DNA can be detected in the picomolar range with a high selectivity and within less than 20 min.
Cui
Liang
L
State Key Laboratory for Physical Chemistry of Solid Surfaces, The Key Laboratory for Chemical Biology of Fujian Province, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
Ke
Guoliang
G
Wang
Chunming
C
Yang
Chaoyong James
CJ
D005090:Exodeoxyribonucleases / Q000737:chemistry / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009696:Nucleic Acids / Q000235:genetics*; D013997:Time Factors
D009696:Nucleic Acids; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III