title: Enzyme-assisted target recycling (EATR) for nucleic acid detection.
abstract: 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.
authors.affiliation: Chemistry Department, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816, USA. yugeras@gmail.com.
authors.forename: Yulia V
authors.initials: YV
authors.lastname: Gerasimova
authors.forename: Dmitry M
authors.initials: DM
authors.lastname: Kolpashchikov
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D012313:RNA / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D015345:Oligonucleotide Probes; D012313:RNA; D004247:DNA; D004262:DNA Restriction Enzymes