The Study On The Establishment Of CiELISA For The Detection Of Furazolidone Residue In Marine Product And The Preparation Of Antiserum To Chloramphenicol

This article can be divided into two parts, one part covers the topic of trying to establish Enzyme-Linked Immunosorbent Assay method for detecting trace element of FZ. The detailed steps will be introduced in the following content. The other part is something about making antiserum to CAP, which will provide the foundation for the further study on establishing an ELISA for detecting CAP in our laboratory.Bovine serum albumin (BSA) and Humane serum albumin (HSA) are used as two protein carriers to couple with the hapten FZ by improved Diazotization procedure. A proper regular immunal program is selected to immunize four healthy New Zealand rabbits to produce pured and high specific antiserum. The indirect competitive ELISA for FZ is established with the antisera. 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 1.25g/ml and 1:3200 relatively. The standard curve of ciELISA is also established and the curve indicats that the lowest detection limit is lug/I 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=95.38l-20.203x (R2=0.9913). The curve has a favorable linearity relation within the concentration range of l~100g/l. However the curve has an unfavorable linearity relation at the concentration exceeding 100g/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 4.16%, lower than 10%.Theintercross ratio of FZ is 9.20%. The average rate of recovery of shirmp muscle by ELISA is 94.86% 9.55%. No cross-reactivity was seen between antibody and five other anitbiotics including chloramphenicol, sulfamethazine, erythromycin, enrofloxacin and trimethoprim. The antibody shows cross-reactivity for furacilinum with no more than forty-three percentage. There are still a great deal of tests needed to do repeatedly by different laboratories and different researchers to further testify the dependability of this established ELISA protocol.