<正>Objective:To optimize the ELISA for the determination of tetrodotoxin.Methods:A corn petitive enzyme-linked immunosorbent assay（ELISA） was used.In the ELISA.100μl antigen（1.0μg/ml） was coated on the microtiter plate for 60 min at 37℃or over night at 4℃.The plate was then washed 3 times with PBS-T for 3-5 s each time.The optimal incubation time for monoclonal antibody （mAb）

goat anti-mice IgG peroxidase conjugate and OPD were 30 min.20 rain and 10 min at 37℃

re- spectively.Results:The detection limit is 0.05 ng in each well.The curve was linear for TTX doses be tween 5 5 000 ng/ml （0.25 250 ng for every assay）.The linear regress equation was Y=0.30 88X-- 0.17 41 （R=0.99 01）.The average callback for TTX of muscles and gonads were 99.74% and 100.30%

respectively.The sensitivity of optimization ELISA was 5 times than traditional method and the time of 1.8 h were saved.Conclusion..The optimized ELISA is an ideal method for the determination of tetrodotoxin.