Identification of protein for the diagnosis of Bartonella henselae infection using an immunoproteomic approach

Abstract

Bartonella henselae, the etiological agent of Cat-scratch disease, is the Bartonella species most commonly detected in cats and humans in Brazil. The approaches currently used for the diagnosis of this disease have limitations in terms of sensitivity or are overly elaborated in its execution and time-consuming. For that matter, immunochromatography is a quick, simple, and less expensive alternative for the diagnosis of bartonellosis caused by B. henselae. The assay does not require intensive training and does not require the use of any equipment; thus, this approach can be easily carried out in veterinary clinics and hospitals by doctors and veterinarians themselves. Thus, in this initial stage, the present study aims to identify and characterize the immunogenic proteins of B. henselae for the use of rapid diagnostic tests. For that purpose, the protein extract of B. henselae will be submitted to electrophoresis (SDS-PAGE), followed by immunoblotting using serum samples from experimentally infected dogs. After the identification of immunoreagent spots, they will be subjected to mass spectrometry to identify the peptides. Subsequently, aiming at the identification of these proteins, the resulting data will be compared with different databases and their characterization will be carried out using various bioinformatics tools. Once identified, the genes responsible for the expression of these proteins will be accessed in the future. The identification of these immunogenic proteins will be the first and primary step for future research, which will assist in the development of new strategies for the diagnosis of bartonellosis in animals and humans. These genes should be cloned in the future and their respective proteins expressed heterologously. Finally, these proteins can be used for the development of immunochromatography assays. Due to the limitations of the diagnostic methods currently used, we believe that a new approach is necessary and would be of great help in the diagnosis of this disease. As there are no alternative methodologies on the Brazilian market to the diagnosis of bartonellosis, the development of this project should result in knowledge that will allow the creation of an efficient product, of quick execution, and accessible cost for the Brazilian market. (AU)