Label-free detection of target DNA sequence and single-base mismatch in hepatitis C virus corresponding to oligonucleotide by resonance light scattering technique

Abstract

A highly sensitive assay to detect sequence-specific DNA by the resonance light scattering (RLS) technique has been developed based on the enhanced RLS intensities of the RLS spectra at 399.5 nm. It was found that Hoechst 33258, when bound to dsDNA and ssDNA in electrolyte solution, displayed different RLS signals. This encouraged us to perform a direct and unlabelled sequence-specific DNA assay. At optimal conditions, this RLS assay can detect complementary target DNA over a range of 4.0 × 10⁻⁹–3.8 × 10⁻⁷ mol L⁻¹, and the limit of detection was around 1.7 × 10⁻⁹ mol L⁻¹. Additionally, the results of experiments showed that different RLS signals reflected a different degree of mismatch between probe DNA and target DNA, and mismatched variants of target DNA with single-base difference can even be well discriminated. Herein, we moved the RLS technique one-step further toward sensitivity, rapidity, simplicity and non-toxicity. The RLS assay results were identified with a fluorescence method, fluorescence polarization assay and atomic force microscope measurement.