Open AccessEntropy Beacon: A Hairpin-Free DNA Amplification Strategy for Efficient Detection of Nucleic Acids
Author(s) -
Yifan Lv,
Liang Cui,
Richard Peng,
Zilong Zhao,
Liping Qiu,
Huapei Chen,
Jinghui Cheng,
Xiaobing Zhang,
Weihong Tan
Publication year - 2015
Publication title -
analytical chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.117
H-Index - 332
eISSN - 1520-6882
pISSN - 0003-2700
DOI - 10.1021/acs.analchem.5b02654
Subject(s) - chemistry , nucleic acid , molecular beacon , thermostability , dna , base pair , entropy (arrow of time) , multiple displacement amplification , detection limit , biophysics , biological system , biochemistry , oligonucleotide , polymerase chain reaction , enzyme , chromatography , gene , thermodynamics , biology , physics , dna extraction
Here, we propose an efficient strategy for enzyme- and hairpin-free nucleic acid detection called an entropy beacon (abbreviated as Ebeacon). Different from previously reported DNA hybridization/displacement-based strategies, Ebeacon is driven forward by increases in the entropy of the system, instead of free energy released from new base-pair formation. Ebeacon shows high sensitivity, with a detection limit of 5 pM target DNA in buffer and 50 pM in cellular homogenate. Ebeacon also benefits from the hairpin-free amplification strategy and zero-background, excellent thermostability from 20 °C to 50 °C, as well as good resistance to complex environments. In particular, based on the huge difference between the breathing rate of a single base pair and two adjacent base pairs, Ebeacon also shows high selectivity toward base mutations, such as substitution, insertion, and deletion and, therefore, is an efficient nucleic acid detection method, comparable to most reported enzyme-free strategies.