earticle

영어

A simple colorimetric method for single nucleotide polymorphisms (SNPs) detection is described using gold nanoparticles (AuNPs) and allele-specific polymerase chain reaction (ASPCR) products. The current strategy is based on the employment of forward primers with thiol group
at 5’ end which after PCR can successfully discriminate the type of target in a simple AuNPs assay. In the case of wild type target DNA, thiolated amplicon is generated by PCR, which upon mixing with AuNPs solution, bind to the surface of nanoparticles (due to the strong gold- thiol interaction) and prevent salt-induced aggregation of particles exhibiting red color to the solution. On the contrary, when the target DNA has single base mutation, PCR cannot be performed, resulting in the absence of thiolated amplicon. Adding this solution to AuNPs followed by NaCl shows blue colored solution suggesting salt-induced aggregation of AuNPs. Applicability of this strategy was successfully demonstrated by detecting single base mutations in breast cancer susceptibility gene BRCA1. This new method enables simple and reliable detection of SNPs without any specialized equipment.