▎ 摘　　要

NOVELTY - Preparing immune sensor involves synthesizing magnetically oxidized graphene by adding graphene material (30-50mL) and graphene oxide (1 mg/mL) in a clean bottle and performing ultrasonic treatment for 1 hour. The product of ultrasonic treatment is obtained in which 0.05-0.07mol/L ferrous chloride solution (20-30mL) and 0.10-0.15mol/L ferric chloride solution (20-30mL) are added at 720-90 degrees C for 3-5 hours, stirred at reflux and 20-30% ammonia solution (10-15mL) is dropped slowly to adjust the pH to 10-11. USE - Method for preparing immune sensor used for detecting pathogenic bacteria of ocean (claimed). ADVANTAGE - The method enables to prepare immune sensor in a simple way and provides fast, sensitive, accurate and highly specific results. DETAILED DESCRIPTION - Preparing immune sensor involves synthesizing magnetically oxidized graphene by adding graphene material (30-50mL) and graphene oxide (1 mg/mL) in a clean bottle and performing ultrasonic treatment for 1 hour. The product of ultrasonic treatment is obtained in which 0.05-0.07mol/L ferrous chloride solution (20-30mL) and 0.10-0.15mol/L ferric chloride solution (20-30mL) are added at 720-90 degrees C for 3-5 hours, stirred at reflux and 20-30% ammonia solution (10-15mL) is dropped slowly to adjust the pH to 10-11. The obtained product is then cooled , washed with water to a pH of 7 and total volume is make up to 50mL to obtain solution of magnetically oxidized graphene material which is then stored at 4 degrees C. The obtained magnetically oxidized graphene solution (200 mu L) is taken in a test tube and kept for ultrasonic treatment for 1 hour. Coupling reagent (200-400 mu L) is added to the obtained solution and dilute hydrochloric acid is added dropwise to adjust pH to 4-6. The solution is then incubated for 1 hour and washed three times with water. The obtained solution is mixed uniformly with 30-50 mu L of electrochemical luminous body solution (0.001-0.1 mu g/mL) and ocean pathogenic bacteria antibody solution (0.1-1 mu g/mL). The aqueous sodium hydroxide solution (0.1mol/L) is added dropwise to the solution to adjust the pH at 8-10, oscillated for 4 hours and washed three times with water. The solution is incubated with 2% bovine serum albumin solution (50-100 mu L) to close non specific active site, at 4 degrees C for 1 hour. The solution is again washed three times with water and volume is make up to 200 mu L to obtain solution of ocean pathogenic bacteria antibody and electrochemical luminous body at the same time marks multifunctional graphene material solution. The obtained product is then stored at 4 degrees C. The light-emitting electrochemical immune sensor is assembled by taking diameter of magnetic glassy carbon electrode in the range of 3-5mm. The electrodes with dimensions of 1.0 mu m, 0.3 mu m, 0.05 mu m are treated with a polishing slurry comprising ethanol and water followed by ultrasonic treatment for 2 minutes in the presence of nitrogen. The product is dried and graphene material solution (5-10 mu L) is added dropwise to the magnetic glassy carbon electrode surface to obtain light emitting electrochemical immune sensor for detecting pathogenic bacteria of ocean. An INDEPENDENT CLAIM is included for a method of detecting marine pathogens by immune sensor, which involves: (A) soaking immune sensor in a solution containing marine bacteria, incubating for 1 hour at 37 degrees C and removing water; (B) preparing three electrode system by taking molybdenum electrode as working electrode, silver or silver chloride as counter electrode and saturated mercurous chloride as reference electrode; (C) dipping the three-electrode system into the buffer solution for initiating electrochemical reaction and measuring the luminous intensity; (D) obtaining different concentrations of marine bacteria solution corresponding to intensity values immune sensor; and (E) establishing quantitative relationship between concentration of pathogens for detecting concentration of sample in marine bacteria.