The development and optimization of ELISA for the determination of tetrodotoxin

• Published in: Journal of Medical Colleges of PLA Vol. 22; no. 6; pp. 347 - 351
• Main Author: 周玉 李岩松 潘风光 柳增善 王哲
• Format: Journal Article
• Language: English
• Published: Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses, Jilin University, Changchun 130062, China%College of Light Industry and Economics & Management, Jilin University, Changchun 130062, China 01.12.2007
• Subjects: 免疫检测法; 单克隆抗体; 最优化; 河豚毒素; monoclonal antibody; tetrodotoxin; ELISA
• ISSN: 1000-1948
• DOI: 10.1016/S1000-1948(08)60016-7

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 C or over night at 4 C. 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, 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.