One-Pot Colorimetric Nucleic Acid Test Mediated by Silver Nanoparticles for DNA Extraction and Detection

Abstract

This study introduces a one-pot colorimetric nucleic acid test (NAT) platform that integrates silver nanoparticle (AgNP)-based DNA isolation and colorimetric detection of bacterial genes. The NAT platform is comprised with purification and reaction units that enable cell lysis, DNA purification, loop-mediated isothermal amplification (LAMP), and colorimetric detection. In the purification unit, polyethyleneimine (PEI)-capped AgNPs were used as cell lysis agents because of their cell-disrupting and antimicrobial properties and were immobilized on a glass fiber membrane for DNA capture and isolation. The reaction unit enabled colorimetric detection of DNA amplicons, achieved by the synthesis of AgNPs on chromatography paper formed via the reduction of silver ions present on the paper, mediated by the use of sodium ascorbate, a reducing agent, present in the LAMP reagent, after the reaction. AgNPs were formed only in the presence of the target amplicons in the positive samples after reaction at 65 °C for 5 min. Bacterial DNA was efficiently extracted using this method, and Enterococcus faecium was detected with a detection limit of 102 CFU/mL. This platform is a promising alternative for rapid and cost-effective nucleic acid testing in resource-limited settings.