title: Quadruplex priming amplification combined with nicking enzyme for diagnostics.
abstract: Quadruplex priming amplification (QPA) is a straightforward assay that allows isothermal amplification of DNA and possesses an intrinsic real-time detection mechanism. QPA can be employed as a diagnostic tool for both linear and exponential signal amplification. The linear QPA, which is less prone to background activity characteristics of exponential systems, suffers from low sensitivity. To increase the sensitivity, here we introduce specific probe molecules that are designed for combined activities of Bst 2.0 polymerase and Nt.BstNBI nicking enzyme. The current assay, which is suitable for single-tube isothermal signal amplification, has increased sensitivity of plain linear QPA by three orders of magnitude to levels of low femtomolar concentration of target molecules.
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Shota
authors.initials: S
authors.lastname: Gogichaishvili
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA; Institute of Biophysics, Ilia State University, Tbilisi 0162, Georgia.
authors.forename: Levan
authors.initials: L
authors.lastname: Lomidze
authors.affiliation: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA; Institute of Biophysics, Ilia State University, Tbilisi 0162, Georgia. Electronic address: kankia.1@osu.edu.
authors.forename: Besik
authors.initials: B
authors.lastname: Kankia
mesh_terms: D001681:Biological Assay / Q000379:methods*; D002623:Chemistry Techniques, Analytical / Q000379:methods* / Q000592:standards; D054856:G-Quadruplexes; D057230:Limit of Detection; D015335:Molecular Probes / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques
chemical_list: D015335:Molecular Probes
keywords: DNA signal amplification; Nicking enzyme; QPA; Quadruplexes