Encoded hydrogel microparticles with universal mismatch-incorporated DNA probes for highly specific multiplex detection of SNPs

• Published in: Talanta (Oxford) 2022-08, Vol.245, p.123480, Article 123480
• Main Authors: Moon, Hyun June, Mun, Seok Joon, Lee, Jun Ho, Roh, Yoon Ho, Lim, Yong Jun, Bong, Ki Wan
• Format: Article
• Language: English
• Subjects: Encoded hydrogel microparticles; Multiplexed detection; Single nucleotide polymorphism; Universal mismatch
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2022.123480

Hydrogel microparticle-based nucleic acid assays are an attractive detection platform based on their multiplexing capabilities with high sensitivity and specificity. A particular area of interest is single-nucleotide polymorphism (SNP) sensing, where multiple SNPs should be identified in a highly reliable yet economical manner. However, hydrogel microparticles leveraging probe-target hybridization as a key mechanism are hampered by small duplex stability differences arising from single base-pair mismatch. We have developed encoded hydrogel microparticles with DNA probes tailored for multiplex SNP detection. Within the DNA probes, we adopt a widely used base analog (5-nitroindole) so that it substitutes one of the base sequences among DNA probes. The effects of the modification of the probes’ structure on SNP sensing has been tested from multiple perspectives, such as specificity, sensitivity, and available assay temperatures at a given ionic strength. We have validated that our hydrogel microparticles exhibit much higher specificity for a single base-pair mismatch with minimal reduction in sensitivity. Our particles can also detect multiple SNPs located in different target strands, which is a significant challenge for conventional particles. [Display omitted] •Universal mismatch probes (UM probes) enhanced the specificity of hydrogel microparticles-based single nucleotide polymorphism (SNP) sensing by 3.5-fold.•UM probes-incorporated hydrogel microparticles accomplished detection sensitivity (limit of detection) of 38 amol.•Encoded hydrogel microparticles incroporated with UM probes enabled one-pot detection of multiple SNPs with high specificity.