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ORIGINAL ARTICLE
Year : 2019  |  Volume : 39  |  Issue : 5  |  Page : 217-222

A monoclonal enzyme-linked immunoassay for the detection of botulinum neurotoxin type E


1 National Defense Medical Center, Institute of Preventive Medicine, Taipei, Taiwan
2 National Defense Medical Center, Institute of Preventive Medicine; Department and Graduate Institute of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan
3 National Defense Medical Center, Institute of Preventive Medicine; National Defense Medical Center, Graduate Institute of Physiology, Taipei, Taiwan

Correspondence Address:
Dr. Cheng-Che Liu
National Defense Medical Center, Graduate Institute of Physiology, Room 6105, 6F, No. 161, Sec. 6, Minchuan E. Road, Taipei 114
Taiwan
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jmedsci.jmedsci_203_18

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Background and Aim: Botulinum neurotoxin Type E (BoNT/E), one of the most lethal toxin known, is the common contamination in fishery products or fish consumption that causes foodborne botulism. It is necessary to establish a sensitive and specific method for the detection of BoNT/E because of its extremely low lethal dose. Methods: In this study, a practical enzyme-linked immunosorbent assay for BoNT/E detection was developed. The assay is based on the sandwich format using monoclonal antibodies of two distinct specificities. An affinity-purified anti-BoNT/E light chain Mab (1E1) is utilized to adsorb BoNT/E from solution, and the second anti-BoNT/E heavy chain C terminus Mab (5E1) conjugated with peroxidase is then used to form sandwich complexes, and peroxidase allows color development and measurement of optical density at 450 nm. Results: Standard curves were linear over the range of 5–100 ng/ml BoNT/E. The limit of detection was below 10 ng/ml in phosphate-buffered saline buffer. The developed BoNT/E assay also showed no cross-reaction to Type A neurotoxin (BoNT/A) and Type B neurotoxin (BoNT/B). Conclusion: Herein, a sensitive and accurate ELISA for BoNT/E detection was presented. It has the potential to utilize in vivo BoNT/E analysis and contamination monitoring.


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