Desenvolvimento e análise de anticorpos policlonais anti-ldh de plasmodium vivax para o diagnóstico de malária

Abstract

Malaria is one of the most serious public health problems in the world, accounting for high morbidity and mortality. The diagnosis remains the most widely used microscopy, however this approach requires adequate infrastructure and highly skilled professionals for the exams, making it unfeasible in areas of difficult access. Tests for the rapid and simple diagnosis of malaria are commercially available, but none of national origin, complicating the deployment by the Unified Health System (SUS). Faced with this problem, this study has as main objective the production of Plasmodium vivax Lactate Dehydrogenase (pLDH), aiming at the development of polyclonal antibodies anti-LDH able to detect the native antigen in blood samples from patients infected with the intention of offering future perspectives for the development of a rapid diagnostic test for malaria. For this, pvLDH protein was produced by recombinant DNA technology in host cells chemically competent. Therefore, it was purified to inoculation into rabbits and mice Balb /c. The response of the inoculated animals as well as the performance of polyclonal antibodies anti-LDH in the recognition of native antigen were assessed by ELISA immunoassays and sandwich system, respectively. The animals showed good antibody titers anti-pLDH after the third inoculation of the recombinant protein pLDH, and the rabbit antibody response than the response of mice with absorbance values at dilution 1/100, 2,600 and 2,100, respectively. Polyclonal antibodies anti-pLDH were able to recognize the native protein pLDH 131 samples to 154 samples positive malaria and incapable of recognizing human isoforms of LDH when evaluated against malaria negative samples, since the 23 negative samples evaluated, all also remained negative during the tests. The proposal idealized in this study proved extremely viable and promising. The results of this study meet expectations and provided future prospects as regards the use of recombinant pvLDH for the development of polyclonal and monoclonal antibodies functional in murine model for use in rapid diagnostic test (RDT) for malaria in attempt to offer alternatives to diagnosis in Brazil, or for use in immunoassay targeting the monitoring of the disease course.