Banca de DEFESA: LARISSA AKASHI

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : LARISSA AKASHI
DATE: 31/08/2023
TIME: 16:00
LOCAL: Sala 306 do Bloco B do Campus de Santo André da Universidade Federal do ABC
TITLE:

Graphene Oxide Conjugated with Trizol Reagent, Magnetic Beads and Iron Nanoparticles for RNA Extraction Methods of Rabies Virus

PAGES: 133
BIG AREA: Ciências Biológicas
AREA: Microbiologia
SUMMARY:

Throughout history, various viruses have stood out as causes of diseases that have plagued
humanity. Rabies, along with smallpox, has been described since ancient times. While rabies still
causes approximately 60,000 human deaths each year, smallpox was eradicated in the 1980s. More
recently, COVID-19, caused by the Severe Acute Respiratory Syndrome Coronavirus type 2 (SARS-
CoV-2), has become a pandemic affecting healthcare systems worldwide. Molecular biology has been
essential in monitoring the evolution of these viruses, with RNA extraction being the initial step for
various methodologies such as Reverse Transcription followed by Polymerase Chain Reaction (RT-
PCR). Laboratory techniques are constantly improving, and the same applies to RT-PCR, hence the
need to search for more efficient and cost-effective tools. With the emergence of nanoscience, carbon
materials such as graphene oxides (GO) have shown efficacy due to their biocompatibility, low cost,
and the manipulation of matter at the molecular level, aiming to create new products applied in
biological processes. In this dissertation, the objective was to incorporate graphene materials into
different RNA extraction methods, primarily for the rabies virus (RABV), but also to evaluate their
potential use with SARS-CoV-2. The GOs were synthesized using the modified Hummers method and
characterized by Raman and UV-Vis spectroscopy with different oxidation times ranging from 1 to 5
hours. They were used in three RNA extraction methodologies: Trizol reagent with GO, commercially
combined with magnetic beads and ferromagnetic nanoparticles synthesized in the laboratory
functionalized with GO (GO-MNPs). The efficiency of RNA extraction with GO was evaluated by
quantifying the RNA obtained in each method using a microvolume spectrophotometer. Subsequently, RT-PCR was performed to confirm the viability of the extracted RNA. It was observed that the interaction between GO and the Trizol reagent was not effective for RNA extraction from RABV. However, methods that used commercially combined magnetic beads with GO and laboratory-synthesized GO-MNPs allowed obtaining higher RNA concentrations, increasing extraction efficiency by 111%. For SARS-CoV-2 extraction, it was also possible to enhance the amount of extracted genetic material and identify better sample purity. The results indicate that the incorporation of GO in RNA extraction can optimize RT-PCR for virus diagnosis, enabling the implementation of more cost-effective molecular methods and improving the efficiency of obtaining extracted nucleic acids.

COMMITTEE MEMBERS:
Presidente - Interno ao Programa - 927.634.300-82 - HELENA BEATRIZ DE CARVALHO RUTHNER BATISTA VICENTE - UFRGS
Membro Titular - Examinador(a) Interno ao Programa - 2605420 - MARIA CRISTINA CARLAN DA SILVA
Membro Titular - Examinador(a) Interno ao Programa - 2605490 - SERGIO DAISHI SASAKI
Membro Titular - Examinador(a) Externo ao Programa - 1764675 - CHRISTIANE BERTACHINI LOMBELLO
Membro Suplente - Examinador(a) Externo ao Programa - 1838179 - KATIA FRANKLIN ALBERTIN TORRES
Membro Suplente - Examinador(a) Externo ao Programa - 3327250 - MÔNICA HELENA MONTEIRO DO NASCIMENTO