Photoinduced atom transfer radical polymerization combined with click chemistry for highly sensitive detection of tobacco mosaic virus RNA.

An electrochemical biosensor for highly sensitive detection of tobacco mosaic virus (TMV) RNA (tRNA) based on click chemistry and photoinduced atom transfer radical polymerization (photoATRP) is developed for the first time. Herein, tRNA is recognized and captured by hairpin DNA immobilized on the gold electrode surface by Au-S self-assembly. Propyl 2-bromoisobutyrate (PBIB), a photoATRP initiator containing an alkyne group, is conjugated to the azide group of hairpin DNA via a Cu(I)-catalyzed azidoalkyl cyclization reaction (CuAAC). Under the irradiation of 470 nm blue light, photoATRP is activated by the photoredox catalyst (eosin Y, EY), resulting in the formation of a large number of electroactive probes (ferrocenylmethyl methacrylate, FMMA), which significantly amplifies the signal. Under the optimal experimental parameters, the strategy has a wide linear detection (0.1 pM-10 nM) (R2 = 0.995) with a limit of detection (LOD) as low as 3.5 fM. In addition, the biosensor also exhibited good selectivity for mismatched bases, excellent stability and reproducibility. Moreover, satisfactory result was achieved when the biosensor was applied to the detection of tRNA from healthy rehmannia total RNA extracts, which demonstrates the great potential of the method in the practical detection of TMV.