Enzyme-assisted target recycling (EATR) for nucleic acid detection
Abbreviated Journal Title
Chem. Soc. Rev.
ULTRASENSITIVE ELECTROCHEMICAL DETECTION; ROLLING CIRCLE AMPLIFICATION; CYCLING PROBE TECHNOLOGY; HYBRIDIZATION CHAIN-REACTION; SEQUENCE-SPECIFIC DETECTION; RESONANCE ENERGY-TRANSFER; DUPLEX-SPECIFIC; NUCLEASE; LABEL-FREE DETECTION; OF-CARE DIAGNOSTICS; ENDONUCLEASE SIGNAL; AMPLIFICATION; Chemistry, Multidisciplinary
Fast, reliable and sensitive methods for nucleic acid detection are of growing practical interest with respect to molecular diagnostics of cancer, infectious and genetic diseases. Currently, PCR-based and other target amplification strategies are most extensively used in practice. At the same time, such assays have limitations that can be overcome by alternative approaches. There is a recent explosion in the design of methods that amplify the signal produced by a nucleic acid target, without changing its copy number. This review aims at systematization and critical analysis of the enzyme-assisted target recycling (EATR) signal amplification technique. The approach uses nucleases to recognize and cleave the probe-target complex. Cleavage reactions produce a detectable signal. The advantages of such techniques are potentially low sensitivity to contamination and lack of the requirement of a thermal cycler. Nucleases used for EATR include sequence-dependent restriction or nicking endonucleases or sequence independent exonuclease III, lambda exonuclease, RNase H, RNase HII, AP endonuclease, duplex-specific nuclease, DNase I, or T7 exonuclease. EATR-based assays are potentially useful for point-of-care diagnostics, single nucleotide polymorphisms genotyping and microRNA analysis. Specificity, limit of detection and the potential impact of EATR strategies on molecular diagnostics are discussed.
Chemical Society Reviews
"Enzyme-assisted target recycling (EATR) for nucleic acid detection" (2014). Faculty Bibliography 2010s. 5358.