Evaluation of the expression of the V595E mutation of the BRAF gene in the Urine of Dogs with Urothelial Bladder Carcinoma by qPCR

• Grant number: 21/00922-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2021
• End date: December 31, 2021
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Pathology
• Principal Investigator: Renee Laufer Amorim
• Grantee: Tatiana Adelino Santos
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Bladder or transitional cell urothelial carcinoma is the most common neoplasm in the urinary tract of dogs. It is a neoplasm considered invasive and can present metastases. The definitive diagnosis occurs late, with limitations regarding the sample collection techniques. Recently the V595E mutation of the BRAF gene was indicated as a diagnostic marker present in urine, detected by Digital PCR, a test with the advantage of non-invasive diagnosis. The purpose of this study is to evaluate the qPCR efficacy for detecting the V595E mutation of the BRAF gene in urine samples of dogs with Urothelial Carcinoma of the Bladder. For this, urine samples will be collected from animals suspected of having urothelial carcinoma, through clinical history and ultrasound examination, in addition to confirmation by histopathological examination by biopsy. THE urine sample will be kept frozen for up to 24 hours after collection. Then, DNA extraction will be carried out and qPCR will be performed for the identification and mutation quantification, confirmed by Sanger sequencing. The proposed project, potential, will generate results that will contribute to improving the diagnosis of dogs with clinical suspicion of bladder urothelial carcinoma, as it is a non-invasive method, accurate, easy to perform, and possibly lower cost. With that, they also maximize the chances of early diagnosis and subsequent therapy. The data will be compiled, tabulated, and evaluated with parametric and non-parametric statistical tests. The project on the agenda will develop a technical product that can be offered to little animals.(AU)