Label-free surface-enhanced Raman spectroscopy detection for tyrosine-methionine-aspartate-aspartate (YMDD)-motif mutants of HBV DNA

Abstract

Hepatitis B is a widespread and potentially life-threatening disease caused by the hepatitis B virus, tyrosine-methionine-aspartate-aspartate (YMDD) - motif mutant is a known HBV variant and responsible for the drug resistance during Long-term lamivudine treatment. In this work, spermine-coated silver nanoparticles were used as surface-enhanced Raman scattering enhancement matrices to detect mutations in single sites of the hepatitis B virus single-stranded DNA. The unique Raman finger-print spectrum of DNA based on spermine-coated silver nanoparticles was detected. And the signals from the phosphate backbone allowed to be an internal standard for standardize the surface-enhanced Raman spectrum of DNA oligonucleotides. The spectral changes were quantitatively related to the composition of adenine in the DNA strand, and the correlation performance reached 0.99. The relative standard deviation of surface-enhanced Raman spectroscopy signal corresponding to the tyrosine-isoleucine-aspartate-aspartate (YIDD), tyrosine-valine-aspartate-aspartate (YVDD), and YMDD was 5.43 %, 7.5 %, and 7.28 %, respectively. The proposed method provides a potential tool for YMDD-motif mutants detection and reference for therapeutic drug monitoring.