Open AccessQuadruplex Priming Amplification at a Range of Human Body Temperature
Author(s) -
David Gvarjaladze,
Tamari Gulua,
Shota Gogichaishvili,
Jozef Hritz,
Nunu Metreveli
Publication year - 2020
Publication title -
biointerface research in applied chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.216
H-Index - 11
ISSN - 2069-5837
DOI - 10.33263/briac111.79327942
Subject(s) - primer (cosmetics) , chemistry , priming (agriculture) , dna , fluorescence , biophysics , biochemistry , biology , organic chemistry , physics , botany , germination , quantum mechanics
Quadruplex priming amplication (QPA) is a very simple amplication assay in which the isothermal amplication is performed using an only DNA polymerase, and detection is conducted by the intrinsic uorescence of the primers. QPA employs specic G-rich sequences as primers that, upon polymerase elongation at specic temperatures, spontaneously dissociate from the primer binding sites (PBS) and fold into a monomolecular quadruplex. Fluorescent nucleotide analogs, such as 3-methyl isoxanthopterin (3MI), when incorporated into these primers, emit light upon a quadruplex formation and permit simple, specic, and sensitive quantication without the attachment of probe molecules. Previously has developed QPA assays with truncated targets and potassium cations that demonstrate an optimal amplication around 55°C. Here, we designed QPA assays with truncated target and led cations at a range of human body temperature. Lead cations reveal the most rapid amplification than potassium and strontium cations. QPA can be applied as a method for the implementation of simple and cheap diagnostics (point of care (POC)), as well as development at a range of 35-45°C temperature, which will make this method more convenient for using it in molecular diagnostics.