A1 Journal article – refereed
Quantitative and discriminative analysis of nucleic acid samples using luminometric nonspecific nanoparticle methods




List of Authors: Pihlasalo S, Mariani L, Harma H
Publisher: ROYAL SOC CHEMISTRY
Publication year: 2016
Journal: Nanoscale
Journal name in source: NANOSCALE
Journal acronym: NANOSCALE
Volume number: 8
Issue number: 11
Number of pages: 10
ISSN: 2040-3364
eISSN: 2040-3372

Abstract


Homogeneous simple assays utilizing luminescence quenching and time-resolved luminescence resonance energy transfer (TR-LRET) were developed for the quantification of nucleic acids without sequence information. Nucleic acids prevent the adsorption of a protein to europium nanoparticles which is detected as a luminescence quenching of europium nanoparticles with a soluble quencher or as a decrease of TR-LRET from europium nanoparticles to the acceptor dye. Contrary to the existing methods based on fluorescent dye binding to nucleic acids, equal sensitivities for both single-(ssDNA) and double-stranded DNA (dsDNA) were measured and a detection limit of 60 pg was calculated for the quenching assay. The average coefficient of variation was 5% for the quenching assay and 8% for the TR-LRET assay. The TR-LRET assay was also combined with a nucleic acid dye selective to dsDNA in a single tube assay to measure the total concentration of DNA and the ratio of ssDNA and dsDNA in the mixture. To our knowledge, such a multiplexed assay is not accomplished with commercially available assays.


Last updated on 2019-20-07 at 17:09