Development of an immunochromatographic test for the diagnosis of feline bartonellosis (Bartonella henselae)

Abstract

Bartonella henselae, the etiologic agent of cat scratch disease (CSD), is the most commonly Bartonella species detected in cats and humans in Brazil. The felines are reservoirs of the bacteria, and the domestic cat is play the main role in the transmission of the pathogen to the human. Studies carried out in the state of São Paulo indicate that until 90% of cats can be infected with B. henselae. So, the identification and correct diagnosis of positive animals is importance for the animal and public health. The approaches currently used for the diagnosis of this disease show limitations in terms of sensitivity or are too elaborate in its execution and is extremely time-consuming. In this way, the immunochromatographic test proves to be an alternative quick, simple and at a lower cost for the diagnosis of bartonellosis caused by B. henselae. Furthermore, such an approach can be easily performed in situ in the veterinary clinics and hospitals by the veterinarians themselves because it does not require intensive training and does not require the use of any equipment. In this scenario, the present study aims to develop an immunochromatography kit for the detection of anti-B. henselae using the p58 recombinant protein. For this, we will perform the cloning gene and subcloning in the pET SUMO vector for the heterologous expression of protein previously identified as immunodominant (phase I results). Due to the limitations of the diagnostic methods currently used, we believe that this new approach is necessary and will be of great value in the aid in the diagnosis of this disease. As they do not exist in the national market alternative methodologies for the diagnosis of bartonellosis, the development of the present proposal will result in the creation of an efficient product, fast and affordable for the national market. In addition to the evaluation of p58 recombinant protein reactivity through immunochromatography, the protein will be used in ELISA and Western-blotting (WB) assays and confronted with serum samples from cats naturally infected with B. henselae (AU)