Development of 1,1′-oxalyldiimidazole chemiluminescent biosensor using the combination of graphene oxide and hairpin aptamer and its application

Highly sensitive biosensor with 1,1′-oxalyldiimidazole chemiluminescence (ODI-CL) detection was developed to rapidly quantify Vibrio (V) parahaemolyticus without time-consuming procedures such as multiple long-incubations and washings. When V. parahaemolyticus in Tris–HCl (pH 7) and hairpin DNA aptamer conjugated with TEX615 in DNA free deionized water were consecutively added in PBS buffer (pH 7.4) containing graphene oxides (GOs), V. parahaemolyticus and GOs bind competitively to hairpin DNA aptamer conjugated with TEX615 during 10 min of incubation at room temperature. Brightness of light immediately emitted with the addition of ODI-CL reagents (e.g., ODI, H2O2) after the incubation was dependent on the concentration of V. parahaemolyticus in a sample. The dynamic range of linear calibration curve for the quantification of V. parahaemolyticus in a sample was from 4375 to 70,000 cells/ml. The limit of detection (LOD=background+3×standard deviation, 2230 cells/ml) of the biosensor operated with good accuracy, precision, and recovery was lower than those of conventional assay methods such as time-consuming and expensive enzyme-linked immunosorbent assays.