Prototyping of a lateral flow assay based on monoclonal antibodies for detection of Bothrops venoms.
- Knudsen C
D Knudsen P
H Laustsen A
Analytica chimica acta[Anal Chim Acta] 2023 Sep 01; Vol. 1272, pp. 341306. Date of Electronic Publication: 2023 May 01.
- Publication Date:
- Background: Brazil is home to a multitude of venomous snakes; perhaps the most medically relevant of which belong to the Bothrops genus. Bothrops spp. are responsible for roughly 70% of all snakebites in Brazil, and envenomings caused by their bites can be treated with three types of antivenom: bothropic antivenom, bothro-lachetic antivenom, and bothro-crotalic antivenom. The choice to administer antivenom depends on the severity of the envenoming, while the choice of antivenom depends on availability and on how certain the treating physician is that the patient was bitten by a bothropic snake. The diagnosis of a bothropic envenoming can be made based on expert identification of the dead snake or a photo thereof or based on a syndromic approach wherein the clinician examines the patient for characteristic manifestations of envenoming. This approach can be very effective but requires staff that has been trained in clinical snakebite management, which, unfortunately, far from all relevant staff has.
Results: In this article, we describe a prototype of the first lateral flow assay (LFA) capable of detecting venoms from Brazilian Bothrops spp. The monoclonal antibodies for the assay were generated using hybridoma technology and screened in sandwich enzyme-linked immunosorbent assays (ELISAs) to identify Bothrops spp.-specific antibody sandwich pairs. The prototype LFA is able to detect venom from several Bothrops spp. The LFA has a limit of detection (LoD) of 9.5 ng/mL in urine, when read with a commercial reader, and a visual LoD of approximately 25 ng/mL.
Significance: The work presented here serves as a proof of concept for a genus-specific venom detection kit that could support physicians in diagnosing Bothrops envenomings. Although further optimisation and testing is needed before the LFA can find clinical use, such a device could aid in decentralising antivenoms in the Brazilian Amazon and help ensure optimal snakebite management for even more victims of this highly neglected disease.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Cecilie Knudsen, Jonas A. Jürgensen, Søren H. Dam, Aleksander M. Haack, Rasmus U. W. Friis, and Andreas H. Laustsen are co-founders of VenomAid Diagnostics A/S. Jonas A. Jürgensen, Pelle D. Knudsen, and Georgina M. Ross are employed by VenomAid Diagnostics A/S. Cecilie Knudsen is an industrial PhD student at the Technical University of Denmark. Her PhD is co-sponsored by Innovation Fund Denmark and BioPorto Diagnostics A/S. Cecilie Knudsen, Jonas A. Jürgensen, Søren H. Dam, Aleksander M. Haack, Rasmus U. W. Friis, and Andreas H. Laustsen have been designated as inventors on a patent application related to the work presented here.
(Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)
- Academic Journal
- Analytica chimica acta
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