| Salmonella belongs to the Enterobacteriaceae families bacteria,which is Gram-negative pathogens,and it is a common food in one of the four major food-borne pathogens,After a lot of experiments,To establish a kind of quickly and accurately Detection methods to provide rapid detection of Salmonella,efficient and convenient way,Immunomagnetic microspheres were synthesized by Salmonella monoclonal antibody conjugated with a Carboxyl magnetic nanosphere,and were used as molecular probes for NMR signal.A novel biosensor for Salmonella based on immune-magnetic beads was prepared by immunology method.And optimizing of process parameters,evaluating of the effectiveness of the new method.The main results are as follows:1.Propagating of Salmonella-specific immune beads and optimizing of the capture rate.2.Optimization of optimum environment of coupling reaction between salmonella and IMBs(the effects of usage of immunomagnetic beads,buffer solution,container and immunocapture time)3.Optimization of Optimal conditions for the solution curve with T2 values.4.Respectively 180°-?-90° pulse sequence and CPMG sequences to measure T2.The T2(Spin-Spin relaxation time)of different samples were investigated by NMR,and the influence of different conditions on the value of T2 were investigated.Meanwhile,the effects of usage of immunomagnetic beads,buffer solution,container and immunocapture time on the sensor response characteristics were studied.Under the optimal conditions,the resulting biosensor displayed a rapid response to Salmonella.The T2 values curve was best when putting 50ug immunomagnetic beads and 1mL PBS(0.01mol/L,pH7.4)into polypropylene centrifuge tube,and mixing for 1.5h.It showed a linear range from 104-107CHU/mL.The novel method was successfully applied for the determination of Salmonella,it would be more short and convenient to detect food-borne pathogenic bacteria in the future,and open up a broad space for low-field NMR technology research and application. |
PDF Full Text Request