Otimização do conjugado Anti-IGG canino no kit diagnóstico da Leishmaniose Visceral Canina (LVC)

Abstract

Introduction: The diagnosis of canine visceral leishmaniasis (CVL) is of great importance in the control of visceral leishmaniasis, a disease that is neglected worldwide and represents a significant public health problem. Enzyme-linked immunosorbent assay (ELISA) test is widely used because of the possibility of large-scale diagnosis and low cost. Objective: The objective of this study was to optimize the stoichiometry of the conjugate used in the EIE/CVL kit; for this purpose, some conjugation and monitoring parameters were modified through molecular exclusion chromatography to improve the final product. Method: The conjugate produced by Bio-Manguinhos was optimized by purification using SEC gel filtration chromatography and evaluated using the EIE-leishmaniasis-visceral-canine-Bio-Manguinhos kit (EIE-LVC). The conjugates were produced with different stoichiometries (IgG X peroxidase): current conjugate, optimized conjugate and optimized and purified conjugate. Results: The results obtained were validated using the manual Canine Visceral Leishmaniasis test. The homogeneity of the conjugate means in the ELISA test was evaluated using the chi-square test, thereby demonstrating the normality and reliability of the data. Based on statistical data, positive samples obtained by the ELISA test showed homogeneity at a significance level of 0.05, i.e. a data reliability level of 95%. The optimization of the production of the canine anti-IgG conjugate, an extremely important component used in the composition of the Bio-Manguinhos EIE LVC KIT, contributed to ensuring the quality of the kit and improving the diagnosis of canine visceral leishmaniasis disease, making it more sensitive and inexpensive for public health. Conclusions: This work demonstrates that it is possible to optimize the anti-IgG canine peroxidase (HRP) conjugate used in the EIE CVL kit to make it more specific and ultimately more economical.