Hairpin probes based click polymerization for label-free electrochemical detection of human T-lymphotropic virus types II.

A novel, simple, and label-free amplification electrochemical impedance method for quantitative detection of Human T-lymphotropic virus types II(HTLV-II) via click chemistry-mediated of hairpin DNA probes (hairpins) with polymers was developed. The hairpins were firstly attached to the gold electrode surface by an S-Au bond, the azido terminals of hairpins were close to the electrode surface, which make it difficult to be approached. After hybridizing with HTLV-II, the hairpins were unfolded and experienced a big configuration change, which made the azido terminals of the hairpins available to conjugate with alkynyl-containing polymer, called P(DEB-DSDA), formed by 1,4-diacetylenebenzene (DEB) and 4,4'-Diazido-2,2'-stilbenedisulfonic acid disodium salt tetrahydrate (DSDA) via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC). With amount of P(DEB-DSDA) conjugated with the hairpin probes via click polymerization, its electrochemical signal can have a great amplification. Under optimized experimental conditions, this new probe showed a low detection limit of 0.171 pM with a good liner in the range of 1 pM-1 nM. Meanwhile, the biosensor also exhibited good selectivity and reliability in detection of real serum samples, indicating that it has great application potential in clinical DNA diagnosis and detection.