Abstract
Botulism is caused by exposure to botulinum neurotoxins (BoNTs). BoNTs are proteins secreted by some species of clostridia; these neurotoxins are known to interfere with nerve impulse transmission, thus causing paralysis. Botulism may be contracted through consumption of food either naturally or intentionally contaminated with BoNT. The human lethal dose of BoNT is not known but is estimated to be between 0.1 and 70 μg; thus, it is important to be able to detect small amounts of this toxin in foods to ensure food safety and to identify the source of an outbreak. Our laboratory previously reported on the development of Endopep-MS, a mass-spectrometric-based endopeptidase method for the detection and differentiation of BoNT. This method can detect BoNT at levels below the historic standard mouse bioassay in clinical samples such as serum, stool, and culture supernatants. We have now expanded this assay to detect BoNT in over 50 foods including representative products that were involved in actual botulism investigations. The foods tested by the Endopep-MS included those with various acidities, viscosities, and fat levels. Dairy and culturally diverse products were also included. This work demonstrates that the Endopep-MS method can be used to detect BoNT/A, /B, /E, and /F in foods at levels spiked below that of the limit of detection of the mouse bioassay. Furthermore, we successfully applied this method to investigate several foods associated with botulism outbreaks.
Highlights
Botulinum neurotoxin (BoNT) is generated by Clostridium botulinum in addition to some other clostridia species; exposure to the toxin causes botulism, a potentially fatal neuroparalytic disease
The Endopep-MS method has been used successfully in the analysis of clinical specimens and culture supernatants for BoNT/A, /B, /E, and /F, demonstrating its potential to diagnose botulism, a potentially fatal disease. This method has limits of detection in clinical specimens and culture supernatants lower than that of the historically standard assay, the mouse bioassay, until now it was uncertain if the assay could be adapted to examine foods directly for the presence of BoNT/A, /B, /E, and /F
Detection of BoNT in foods is important as it allows for discovery of the vehicle of transmission in foodborne botulism cases, potentially preventing additional outbreaks
Summary
Botulinum neurotoxin (BoNT) is generated by Clostridium botulinum in addition to some other clostridia species; exposure to the toxin causes botulism, a potentially fatal neuroparalytic disease. The use of an immunoaffinity step prior to incubation with the peptide substrate has proven effective at detecting and differentiating BoNT in clinical specimens[19] and culture supernatants.[20] The Endopep-MS method achieves limits of detection comparable to or below that of the historically used mouse bioassay.[21]
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