Patent ID: 8951403
Filing Date: 2015-02-10
Classification: C12Q,G01N

Abstract:
1. A detection method comprising: (1) adding 1.0 g of N-isopropylacrylamide (NIPAAm) powder to 20 ml water, heating at 60° C. to form a NIPAAm solution, and cooling the NIPAAm solution to room temperature; (2) adding 200 μl of 98.7 wt % of ammonium peroxodisulfate (APS) and 50 μl of 99 wt % of N,N,N,N-tetramethylethylenediamine (TEMED) to the NIPAAm solution, uniformly mixing it to form a mixture, and reacting the mixture for 30 hours at room temperature to prepare a transparent gel, poly-N-isopropylacrylamide (PNIPAAm); (3) adding 5 mg enzymes to 100 μl of 1×PBS buffer solution, uniformly mixing it, adding 100 μl of the transparent gel, PNIPAAm to the buffer solution, and uniformly mixing the buffer solution; (4) placing a biosensor on a heater for heating at a constant temperature of 37° C., wherein the biosensor is an Electrolyte-insulator-Semiconductor (EIS) sensor having a sensor membrane formed of europium titanium oxide; (5) dropping 25 μl of the buffer solution on the sensor membrane of the biosensor for wrapping the enzymes on the sensor membrane of the biosensor; (6) heating and curing the wrapped biosensor to form a white gel on the sensor membrane; (7) placing the biosensor in a solution kept at 37° C. and keeping the white gel obtained at step (6) at 37° C.; and (8) taking a measurement.