Development of a sensitive enzyme immunoassay (ELISA) for specific identification of Lachesis acrochorda venom

Núñez Rangel V (1, 2), Fernández Culma M (1), Rey-Suárez P (1), Pereañez JA (1, 3)

(1) Program of Ophidism and Scorpionism, University of Antioquia, Medellin, Colombia; (2) School of Microbiology, University of Antioquia, Medellin, Colombia; (3) School of Pharmaceutical Chemistry, University of Antioquia, Medellin, Colombia.

Abstract: The snake genus Lachesis provokes 2 to 3% of snakebites in Colombia every year. Two Lachesis species, L. acrochorda andL. muta, share habitats with snakes from another genus, namely Bothrops asper andB. atrox. Lachesis venom causes systemic and local effects such as swelling, hemorrhaging, myonecrosis, hemostatic disorders and nephrotoxic symptoms similar to those induced by Bothrops, Portidium and Bothriechis bites. Bothrops antivenoms neutralize a variety of Lachesis venom toxins. However, these products are unable to avoid coagulation problems provoked by Lachesis snakebites. Thus, it is important to ascertain whether the envenomation was caused by a Bothrops orLachesis snake. The present study found enzyme linked immunosorbent assay (ELISA) efficient for detecting Lachesis acrochorda venom in a concentration range of 3.9 to 1000 ng/mL, which did not show a cross-reaction with Bothrops, Portidium, Botriechis and Crotalus venoms. Furthermore, one fraction of L. acrochorda venom that did not show crossreactivity with B. asper venom was isolated using the same ELISA antibodies; some of its proteins were identified including one Gal-specific lectin and one metalloproteinase. This test may be useful to physicians, since it could be applicable for tracking the kinetic distribution of antigens in patients or experimentally envenomed animals.

Key words: snake venom, Lachesis, Lachesis acrochorda, enzyme-linked immunosorbent assay.

ACKNOWLEDGEMENTS

The authors thank Prof. Bruno Lomonte (Instituto Clodomiro Picado, Costa Rica) for identification of peaks of Lachesis acrochorda venom (LaSF).

COPYRIGHT

© CEVAP 2012

SUBMISSION STATUS

Received: September 8, 2011.

Accepted: December 20, 2011.

Abstract published online: January 25, 2012.

Full paper published online: May 31, 2012.

CONFLICTS OF INTEREST

The authors declare that there are no conflicts of interest.

FINANCIAL SOURCE

Universidad de Antioquia (CODI project), and COLCIENCIAS (project 1115-459-21441 and Programa de Jóvenes Investigadores) provided the financial grants for this project.

ETHICS COMMITTEE APPROVAL

The present study was approved by the Ethics Committee of Antioquia University, Medellin, Colombia.

CORRESPONDENCE TO

Vitelbina Nuñez Rangel, Programa de Ofidismo/Escorpionismo, Universidad de Antioquia, Medellín, Colombia. Phone: 574 219 65 35. Email: vitelbina.nunez@siu.udea.edu.co.