Development of a rapid thrombophilia test

• Grant number: 19/23088-7
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: August 01, 2020
• End date: April 30, 2021
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Debora Colombi
• Grantee: Debora Colombi
• Company: Genotyping Laboratório de Biotecnologia Ltda (Genotyping)
• CNAE: Testes e análises técnicas ; Pesquisa e desenvolvimento experimental em ciências físicas e naturais ; Atividades profissionais, científicas e técnicas não especificadas anteriormente
• Associated researchers: Alexandre Hideaki Takara ; Lucas Guerchi Belo
• Associated scholarship(s): 20/11420-4 - Development of a rapid thrombophilia test, BP.TT

Abstract

Thrombophilia is a condition that causes changes in blood coagulation, with a consequent increased risk of blood vessel obstruction. Thrombophilia may occur due to mutations or deficiencies in the production of clotting factors and may be hereditary or acquired. Hereditary thrombophilias constitute a heterogeneous group of changes in blood coagulation, which increase the predisposition to the development of thromboembolic phenomena and generally manifest themselves in the presence of other risk factors. Through genetic examination it is possible to know if the person has a genetic (hereditary) predisposition to thrombosis and it is possible for the doctor to take preventive measures. The most common mutations are as follows: Leiden Factor V (R506Q), Prothrombin (G20210A) and MTHFR (677C> T and 1298A> C) mutation. Oral contraceptive use is associated with an approximately 4-fold increase in the risk of thrombosis; In combination with an inherited risk factor the risk rises to 34 times in heterozygotes and more than 100 times in homozygotes. This project aims to develop a test that is cheaper than Sanger Sequencing, the current technique used in the laboratory for this test, and that is faster and easier, thus not requiring very qualified personnel to perform it. For this purpose this project aims to standardize two different technologies: 1) isothermal PCR reaction (DNA amplification) without the need for DNA extraction and with the result by colorimetric visualization, that is, we will associate the use of PCR from raw samples. With the G-quadruplex technique and 2) TETRA-ARMS-PCR and then its comparison with current technology. By the end of the project, Genotyping will have acquired the know-how to develop other theses in the ANS roll with this technology and be able to offer it to other laboratories as a product, instead of just continuing to offer service. (AU)