An Approach to Identifying Single-Nucleotide Mutations Using Noncovalent Associates of Gold Nanoparticles with...

Globally, widespread tuberculosis is one of the acute problems of healthcare. Drug-resistant forms of tuberculosis require a personalized approach to treatment. Currently, rapid methods for detecting drug resistance of Mycobacterium tuberculosis (MTB) to some antituberculosis drugs are often used and involve optical, electrochemical, or PCR-based assays. Despite the large number of these assays, it is necessary to develop new tests (for drug-resistant MTB strains) that are structurally simple and do not require specialized equipment. Colorimetric assays involving a colloidal solution of gold nanoparticles (AuNPs) have good potential for the development of the needed diagnostic tools. Here, conditions were found for the formation of tandem duplexes between DNA probes and DNA targets, representing a part of MTB gene gyrA, either wildtype or containing a single-nucleotide polymorphism associated with fluoroquinolone resistance of MTB. Adsorption of the duplexes on AuNPs allowed to distinguish the two targets owing to the formation of nano-constructs of different structures. Interaction of DNA with AuNPs was analyzed by optical spectroscopy, dynamic light scattering, and transmission electron microscopy. A scheme is proposed for direct colorimetric detection of the fluoroquinolone-resistance-associated single-nucleotide polymorphism at a 2 nM concentration in a liquid system based on a shift of AuNPs' optical absorption maximum.