The Study On The Establishment Of CiELISA For The Detection Of Sulfamethazine Residue In Aquatic Products And Rudimental Apply

This article can be divided into two parts, In the first part,Bovine serum albumin(BSA) and Human serum albumin(HSA) were used as two protein carriers respectively to couple with the hapten Sulfamethazine(SM2) by improved Diazotization procedure.The complete antigens SM2 - BSA and SM2 - HSA were acted as immunoantigen and coating antigen respectively in ciELISA. Selected a few of immunal programs to immunize the New Zealand rabbits to produce pured and high specific antiserum. The second part covers the topic of trying to establish the indirect competitive Enzyme-Linked Immunosorbent Assay method for detecting trace element of SM2. The most suitable working condition of ciELISA is obtained by several grid tests which are used to calculate the most optimal concentration and dilution of the coating antigen and antiserum, and the result is l.0μg/ml and 1:3200 relatively. The standard curve of ciELISA is also established and the curve indicats that the lowest detection limit is 1.89μg/L,which can provide some valuable datas used for the demanded detection limit of ND(No-Detetion) at home and abroad. The regression equation of the curve is y=88.323 — 18.013x, (R2=0.9964) The curve has a favorable linearity relation within the concentration range of 10~200μg/L .However the curve has an unfavorable linearity relation at the concentration exceeding 200μg/L.The accuracy of the established ELISA protocol can be determined by errors within one batch and among batches. The error within one batch is 2.56%, lower than 10%.The intercross ratio of SM2 is 5.14%. The average rate of recovery of Litopenaeus vannamei muscle and Paralichthys olivaceus muscle by ciELISA is 90.98%±3.32% and 90.02% ±2.68%. No cross-reactivity was seen between antibody and five other anitbiotics including Sulfamethoxazole, Chloramphenicol, Enrofloxacin, Trimethoprimand Furazolidone. The antibody shows cross-reactivity for Sulfadiazine with no more than eleven percentage.It showed that this is a method with high specificity and selectivity,which can detect the residue Sulfamethazine quickly.