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EVALUATION OF THE QUANTITY AND QUALITY OF DNA AND mRNA EXTRACTED FROM PARAFFIN BLOCKS OF MAST CELL TUMOR WITH DIFFERENT FIXATION TIMES

Grant number: 23/03366-8
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Start date: July 01, 2023
End date: June 30, 2024
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Principal Investigator:Carlos Eduardo Fonseca Alves
Grantee:Leonardo Augusto Moreira Santos
Host Institution: Vice-Reitoria de Pesquisa e Pós-Graduação. Universidade Paulista (UNIP). São Paulo , SP, Brazil

Abstract

With the development of veterinary medicine, new diagnostic and therapeutic techniques emerged, providing a greater animal life expectancy. As a result, veterinary medicine is experiencing an increasing frequency of age-related diseases such as cancer. Along with this increase, there was a growing interest in veterinary oncology, as well as an increase in the number of researches focused on the standardization of molecular tests from the paraffin samples of these patients. Among these tests immunohistochemistry, qPCR and sequencing from tissue tests can be highlighted as having a considerable increase in routine usage. However, for these techniques to be used with a translational aspect, they need to adapt to the clinical routine of veterinary oncology. And with that, most samples available for evaluation in canine patients are formalin-fixed and paraffin-embedded (FFPE). With the advancement of these techniques in human oncology, several studies have evaluated the quality, quantity and applicability of paraffin material for different molecular techniques in oncological diagnosis. However, for veterinary medicine, studies are focused on immunohistochemistry. Several veterinary works have already evaluated the fixation times in relation to the interference in the immunohistochemistry result, and today it is a standard that the sample stays for 48 to 72 hours in the fixative until processing. However, for molecular analyses, there are no specific researches that make this assessment in dog specimens. Human studies have shown that there is a great interference in the fixation time related to the possibility of performing these exams, although, no study has determined the effect of fixation on quantity and quality of genetic molecules in veterinary patient samples. With the growth of molecular tests using paraffin-embedded samples in veterinary oncology, it is necessary to establish the effect of fixation time on the quality of biological samples. In this context, this research aims to evaluate the quality of DNA and mRNA subjected to different fixation times.

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