Ultrasensitive fluorescent detection of HTLV-II DNA based on magnetic nanoparticles and atom transfer radical polymerization signal amplification

Abstract

Human T-lymphotropic virus type II (HTLV-II) is a crucial retrovirus that is closely associated with a variety of human diseases. Herein, an ultrasensitive fluorescent HTLV-II DNA detection strategy was developed for the first time based on magnetic nanoparticles (MNPs) and atom transfer radical polymerization (ATRP) amplification. In this approach, hairpin DNA probes (pDNA) labelled with 5′ thiol and 3′ azide group terminally were immobilized on amino group modified MNPs surface through sulfo-N-succinimidyl-4-maleimidobutyrate sodium salt (sulfo-GMBS) cross-linkers. In the presence of target DNAs (tDNA), pDNA hybridized with tDNA to form double-stranded DNA, and therefore its azide group was away from the MNPs surface. Subsequently, to initiate ATRP reaction, initiators were introduced into the pDNA by a Cu (I)-catalyzed alkyne-azide cycloaddition (CuAAC). Then, large numbers of 9-anthracenylmethyl methacrylate polymer (pAMMA) were successfully labelled on the MNPs surface, resulting in significant amplification of the fluorescence signal. Under optimized conditions, the fluorescence signal was proportional to the logarithm of the concentration of tDNA over the range from 1 fM to 1 nM, with a detection limit of 0.22 fM. Moreover, this strategy was capable of discriminating mismatched bases and detecting HTLV-II DNA in human serum samples. By virtue of the high sensitivity, selectivity, simplicity and economy, this ultrasensitive biosensor demonstrates great potential for biomedical research and early clinical diagnosis.