首页|The development and optimization of ELISA for the determination of tetrodotoxin
The development and optimization of ELISA for the determination of tetrodotoxin
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NETL
NSTL
Objective: To optimize the ELISA for the determination of tetrodotoxin. Methods: A competitive enzyme-linked immunosorbent assay (ELISA) was used. In the ELISA, 100 μl antigen (1.0 μg/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 min and 10 min at 37 C , respectively. Results: The detection limit is 0. 05 ng in each well. The curve was linear for TTX doses between 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.
NETL
NSTL
