Can anti-bothropstoxin-I antibodies discriminate between Bothrops jararaca and Bothrops jararacussu venoms?

Ricardo Teixeira Araujo,Russolina Benedeta Zingali,Carlos Corrêa-Netto,Irene Fernandes,Leonora Brazil-Más,Caio Raony Farina Silveira

doi:10.1186/s40409-017-0105-z

Abstract

BackgroundSnakes of the genus Bothrops, popularly known as pit vipers, are responsible for most cases of snakebite in Brazil. Within this genus, Bothrops jararacussu and B. jararaca deserve special attention due to the severity of their bites and for inhabiting densely populated areas. Regarding the treatment of snakebites by Bothrops jararacussu, questions have been raised about the effectiveness of the specific bothropic antivenom in neutralizing myotoxic effects; however, there are no accurate data for humans. Thus, the development of a differential diagnostic kit for this species would be of great interest because it provides, for healthcare professionals, a tool that would allow us to determine whether the accident was caused by B. jararacussu or other species of the genus. It would also make it possible to evaluate the specificity of the treatment and to provide data for epidemiological studies.MethodsFirst, we produced a species-specific polyclonal antibody – a potential biomarker of Bothrops jararacussu venom – against bothropstoxin-I (BthTx-I), which is also found in smaller quantities in the venoms of B. jararaca from southern Brazil.ResultsPolyclonal antibodies against bothropstoxin-I could be separated into several species-specific immunoglobulins. Then, aiming to develop a system of safe and standardized immunoassay, we produced monoclonal antibodies. Seven hybridomas were obtained. Five of them were specific to the venom of B. jararacussu and two recognized the venom of B. jararaca from the southeastern population. The use of monoclonal antibodies also made it possible to differentiate B. jararacussu from B. jararaca venom obtained from the southern population. Analyzing the reactivity of monoclonal antibodies against other bothropic venoms, we found mAb Bt-3 to be more specific than others for B. jararacussu venom.ConclusionsThese results show the potential of BthTx-I for producing monoclonal antibodies that differentiate between B. jararacussu and other Bothrops species venoms.

Highlights

Snakes of the genus Bothrops, popularly known as pit vipers, are responsible for most cases of snakebite in Brazil
Polyclonal species-specific antibodies We first tested whether the anti-BthTx-I polyclonal antibodies could give specific antibodies to recognize venoms from B. jararacussu and B. jararaca
Analyzing the reactivity of monoclonal antibodies (mAbs) against the venom of B. jararaca from the south of Brazil As soon as we discovered the presence of BthTx-I in the venom of B. jararaca from the south region of Brazil, we evaluated its reactivity to mAbs using ELISA [15]

Summary

Introduction

Snakes of the genus Bothrops, popularly known as pit vipers, are responsible for most cases of snakebite in Brazil Within this genus, Bothrops jararacussu and B. jararaca deserve special attention due to the severity of their bites and for inhabiting densely populated areas. The question of the efficiency of specific bothropic antivenom against B. jararacussu bites is often discussed, since the myotoxic effects observed in mice are not completely neutralized. For this reason, some researchers have suggested the use of a combinated bothropic-crotalic antivenom as a more appropriate treatment [1, 6,7,8,9,10]

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