High-throughput Rapid Detection Method Of Antibiotic In Water

Antibiotics have been widely used in various aspects of human life,which leads to the frequent detection of multiple antibiotics in the water environment and seriously threatens ecological safety and human life and health,.Based on the principle of specificity between antigen and antibody,this study combined with the planar waveguide immunosensor developed by the research group.the method of antibiotic elisa detection and planar waveguide immune sensor detection were established to provide technical support for the monitoring and managementof antibiotics in water environment.Based on the principle of enzyme-linked immunosorbent assay?ELISA?,the optimal coating concentration and antibody concentration of Sulfameter and Ciprofloxacin indirect competitive ELISA were determined by experiments.The blocking time was determined to be at least 100min,the reaction time was at least80min,and the dilution ratio of enzyme-labeled secondary antibody was 1:500.An indirect competitive ELISA assay for Sulfameterand Ciprofloxacin was established.The detection limit of Sulfameter was 17?g/L.The quantitative detection interval was 35.91³61.12?g/L;the detection limit of ciprofloxacin was 1.95?g/L,and the quantitative detection interval was 4.08⁵3.93?g/L.Based on the planar waveguide immunosensor developed by the research group,the preparation of immunochip and the fluorescent labeling process of antibody were explored.The effects of antibody concentration,pH,water hardness and humic acid concentration on the detection of immunosensor were established.The results showed that the concentration of Sulfameter antibody and Ciprofloxacin antibody was 1.5 ug/mL and 1 ug/mL respectively.When pH was 6-8,the concentration of Ca²⁺in water was less than 200 mg/L and humic acid in water was less than 25mg/L,the immune sensor detection was less affected.The detection method of Sulfameter and ciprofloxacin planar waveguide immunosensor was established,and specific experiments and repetitive experiments were carried out.The detection limit of sulfamethoxazole was 0.0052?g/L,and the quantitative detection interval was0.03¹.37?g/L;the detection limit of ciprofloxacin was 0.020?g/L,and the quantitative detection interval was 0.10⁸.83?g/L.Finally,based on the established indirect competitive ELISA method and the planar waveguide immunosensor detection method,the Sulfameter and Ciprofloxacin were respectively spiked and recovered in the actual samples.The recoveries were 80%Between and 115%,the relative standard deviations are less than 15%.The established detection methods have good accuracy,precision and stability,indicating that the established detection methods are reliable.