Associating and Dissociating Nanodimer Analysis for Quantifying Ultrasmall Amounts of DNA

The amplification- and enzyme-free quantification of DNA at ultralow concentrations, on the order of 10-1000 targets, is highly beneficial but extremely challenging. To address this challenge, true detection signals must be reliably discriminated from false or noise signals. Herein, we describe the development of associating and dissociating nanodimer analysis (ADNA) as a method that enables a maximum number of detection signals to be collected from true target-binding events while keeping nonspecific signals at a minimum level. In the ADNA assay for ultralow target concentrations, Au nanoprobes on a lipid micropattern were monitored and analyzed in situ, and newly defined dissociating dimers, which are eventually decoupled into monomers again, were incorporated into the detection results. Tens to thousands of DNA copies can be reliably quantified with excellent single-base-mismatch differentiation capability by this non-enzymatic, amplification-free ADNA method.