Surface Plasmon Resoance Biosensor For Rapid Detection Of Aquatic Pathogens

In recent years,the aquaculture industry in China has developed rapidly and has become a major aquaculture producer in the world.However,due to the high density of intensive culture methods,which make the occurrence of aquaculture diseases more and more frequent,and the impact of bacteria on aquaculture is particularly serious,so the establishment of high sensitivity,good accuracy,rapid detection method is particularly important.The surface plasmon resonance(SPR)biosensor technology has been widely used in food,medicine,biology and other fields as a rapid and efficient detection method.This topic has successfully established SPR biosensor technology for the detection of aquatic pathogens in order to expand the application scope of SPR,laying the foundation for early diagnosis and timely defense of aquatic diseases.The main research contents of this topic are as follows:(1)The antibody was coupled to CM5 chip to establish direct SPR method for direct detection of Aeromonas hydrophila B11.The p H of the coupling antibody buffer,antibody concentration and regenerative liquid were optimized,and the parameters of repeatability,specificity and sensitivity were evaluated.The detection limit of Aeromonas hydrophila was 4.6×10~4 cfu/m L.It has good repeatability and specificity,and can also be used for the detection of real samples.(2)In order to increase the detection sensitivity,a double antibody sandwich method was established for the detection of Aeromonas hydrophila B11.The injection concentration,time,and flow rate of the polyclonal antibody were optimized.The bacteria were tested and evaluated for parameters such as repeatability,specificity and sensitivity.The detection limit of Aeromonas hydrophila B11 was 6.3×10~3 cfu/m L.It has good repeatability and specificity,and can also be used for the detection of real samples.(3)In order to detect different pathogens,we established a dual-channel method for the detection of Aeromonas hydrophila B11 and Edwardsiella B79.The antibodies of Aeromonas hydrophila and Edwardsiella were coupled in two channels,and two bacteria mixed solutions were detected.The detection of the mixture showed that the detection limits of Aeromonas hydrophila B11 and Edwardsiella B79 were 6.6×10~4 cfu/m L and 1×10~5 cfu/m L.It has good repeatability and specificity,and can also be used for the detection of real samples.