Innovation into ofidic accidents: the proof of concept for the development of a quick diagnostic kit

• Grant number: 18/15446-8
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: July 01, 2019
• End date: May 31, 2020
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Laudicéia Alves de Oliveira
• Grantee: Laudicéia Alves de Oliveira
• Company: Paulo Daniel Leite Pesquisa e Desenvolvimento
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais ; Atividades de serviços de complementação diagnóstica e terapêutica
• City: Botucatu
• Associated researchers: Benedito Barraviera ; Lucilene Delazari dos Santos ; Rui Seabra Ferreira Junior
• Associated scholarship(s): 19/14647-2 - USE OF MASS SPECTROMETRY IN THE IDENTIFICATION AND CHARACTERIZATION OF ANTIBODIES FOR PRODUCTION OF A QUICK DIAGNOSTIC KIT, BP.TT; 19/14690-5 - Immunoaffinity chromatography as strategy as a strategy for obtaining specific antibodies antivenom., BP.TT; 19/14668-0 - Imunoenzimatic tests in the standardization of the development of a quick diagnostic kit used in the optimization of the diagnosis of serpent accidents, BP.TT

Abstract

In 2017 the World Health Organization (WHO) reintroduced ophidian accidents into the list of neglected tropical diseases. This decision was based on the fact that annually more than 100,000 people die and more than 400,000 surviving remain with permanent sequelae. In Brazil, in 2017, the SINAN received 224,590 notifications of accidents involving venomous animals with 439 deaths. Of these 30,367 were caused by venomous snakes with 111 deaths. Among the causes of accidents are: 71.41% Bothrops, 7.03% Crotalus, 3.0% Lachesis and 0.78% Micrurus. Most of the affected do not know the aggressor animal, or they do not bring it to the medical consultation, making difficult the correct diagnosis by the health professional. To make matters worse, many of the Brazilian Medical Schools do not teach the theme "accidents for venomous animals" to their students. On the other hand, to date, there is no readily available diagnostic laboratory device that can be used in the hospital emergency room. In this case, the doctor uses the clinical diagnosis to treat the patient. It occurs that it is not possible to differentiate the bothropic accidents from the laquectic, that overlaps in the Amazon region and kills Atlântica; nor the crotalic ones of the elapidic, since the clinical pictures are very similar. In the first case, the doctor will treat the injured individual with bothropic-laquetic serum, when available. In the second case, either the development of the symptoms and the results of the subsidiary laboratory tests are awaited, or the two specific sera, namely, crotalic and elapidic, are treated. This behavior, in spite of being a savior for the injured individual, greatly increases the Unified Health System (SUS), exclusive distributor and sponsor of these antivenoms for all Brazil. In view of this scenario, the technological development of a rapid diagnostic device is relevant for the country because it will facilitate the diagnosis of the snake genus causing the accident, thus allowing the correct diagnosis of the etiological agent, thus indicating the specific treatment. In the present proposal of innovative research for the development of this device, it is proposed to isolate specific antibodies from the antibothropic, anticrotalic, antielapidic and antilaquetic sera produced by the seroproductive Institutes against the venom of the snakes Bothrops jararaca, Crotalus durissus terrificus, Micrurus frontalis and Lachesis muta muta. This experimental stage will be achieved through the development of chromatographic platforms using the venoms of the respective snakes as specific binders. Once these antibodies have been isolated, solid phase immunoenzymatic assays (ELISAs) will be performed using the blood of snake injured individuals before, during and after treatment with the specific sera in order to quantify the level of circulating venom and to evaluate the cross reactivity between these antibodies against the reference venoms. Quality control will be carried out by means of mass spectrometry analyzes. At the end of this proof-of-concept proposal, it is expected that the results obtained will provide technical-scientific data regarding thresholds of sensitivity and specificity, promote the development of a portable device to distinguish between different Brazilian ophidian accidents. This diagnostic device is unprecedented in the national market and will not require electrical energy, battery or chemical reagents and neither refrigerated storage. Given that the technology that will be developed can be transferred internationally, this proposal will have a positive financial impact on the company that intends to constitute, against the data of notifications of ophidian accidents in Brazil and in the world, annually. (AU)