Sensitive biosensor for p53 DNA sequence based on the photothermal effect of gold nanoparticles and the signal amplification of locked nucleic acid functionalized DNA walkers using a thermometer as readout

A convenient photothermal biosensor was constructed for p53 DNA sequence detection based on the high discrimination capability of locked nucleic acid and high efficiency of signal amplification strategy of DNA walkers and difference photothermal effect between aggregated and dispersed gold nanoparticles (AuNPs). The presence of target activated the DNA walkers via the high affinity between target and complementary locked nucleic acid in the probe strand, resulting in the hybridization of the walker strand and substrate strand to form a specific enzyme recognition site. Under the cleavage of the endonuclease, single-stranded DNA (ssDNA) was released to the solution. Then the walker strand bound to a new substrate strand, and the next round of cleavage was triggered. The released ssDNA enhanced the stability of AuNPs against salt-induced aggregation. Given difference photothermal effects of the aggregated AuNPs and dispersed AuNPs under the near-infrared laser, the change of the temperature was detected by a common thermometer easily, which had a linear relationship with the target concentration in the range of 2.0-120.0 pM, the detection limit was 1.4 pM (S/N 1/4 3). The proposed photothermal assay has been applied to detect p53 DNA sequence spiked complex samples with satisfying results.