ELISA Based Detection of Botulinum Neurotoxin Type F in Red Meat and Canned Fish

Abstract

Botulinum neurotoxin (BoNT) is the most toxic protein molecule so far known to animals and humans, due to its extreme toxicity, CDC has been listed as category ‘A' Biological warfare agents. In India, there is no commercial detection system available for the detection of botulism. The present study was aimed to develop an ELISA based detection system for botulinum neurotoxin serotype F in red meat and canned fish. Polyclonal antibodies were generated against the recombinant BoNT/F LC protein and IgG was purified. Sandwich ELISA system was successfully optimized with LOD of ~ 7.8 ng/ml in PBS. Similarly, different concentrations of BoNT/F were spiked in red meat and canned fish and extracted the toxin and the detection LOD was achieved for red meat ~ 62 ng/ml and for canned fish ~ 31 ng/ml. The developed detection system is highly specific. The developed assay will be useful for the screening of botulinum toxins in a large number of food samples.

About Clostridium botulinum, Fish and Tilapia

“…Other alternative or complementary methods to the detection of C. botulinum and toxins in food, unlike the physiological and biochemical characteristics of the culture and the mouse bioassay, the latter considered the gold standard method for detecting botulism, are the Immunological Essay Linked to Enzymes (ELISA), lateral flow immunoassays, electrochemiluminescence (ECL), endopeptidase assays, MALDI TOF mass spectrometry, Polymerase Chain Reaction (PCR), among others (Drevinek et al, 2012;Shahcheraghi et al, 2013;Solomon & Timothy, 2017;Lopez-Villar, 2019;Yadav et al, 2019). For C. botulinum molecular identification is carried out through the Polymerase Chain Reaction (PCR) in its multiple variants and can be used in conjunction with mouse bioassays.…”