Ni-Pt nanozyme-mediated relaxation and colorimetric sensor for dual-modality detection of norovirus.

An NiPt nanozyme-mediated relaxation and colorimetric sensor is developed for dual-modality detection of norovirus (NoV). The relaxation modality is based on the "catalase-like" activity of the NiPt nanozyme, which adjusts the hydrogen peroxide (H2 O2 ) mediated Fe (II)/Fe(III) conversion, thereby changing the relaxation signal. Poly-γ-glutamic acid (MW ≈ 1w) can enhance the relaxivity of Fe(III) (r1  = 7.11 mM-1  s-1 ; r2  = 8.94 mM-1  s-1 ). The colorimetric modality exploits the "peroxidase-like" activity of the NiPt nanozyme, which can catalyze the oxidation of colorless 3, 3', 5, 5'-tetramethylbenzidine (TMB) to blue oxTMB in H2 O2 . Under optimal conditions, the relaxation modality exhibits a wide working range (1.0 × 101 -1.0 × 104 fM) and a limit of detection (LOD) of 4.7 fM (equivalent to 2820 copies/μL). The spiked recoveries range from 99.593 to 106.442 %, and the relative standard deviation (RSD) is less than 5.124 %. The colorimetric modality exhibited the same working range with a lower LOD of 2.9 fM (equivalent to 1740 copies/μL) and an RSD of less than 2.611 %. Additionally, the recombinase polymerase amplification reaction enabled the detection of low NoV levels in food samples with a working range of 102 -106 copies/mL and LOD of 102 copies/mL. The accuracy of the sensor in the analysis of spiked samples is consistent with the gold standard method (real-time quantitative reverse transcription-polymerase chain reaction), demonstrating the high accuracy and practical utility of the sensor.