Banca de DEFESA: Leonardo Ferreira da Silva

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : Leonardo Ferreira da Silva
DATE: 18/12/2023
TIME: 14:30
LOCAL: Auditório 4 do Instituto de Biologia
TITLE:

"Method for detecting SARS-CoV-2 viral RNA using Ribozymes and associated DNA hairpins in hybridization chain reaction with fluorescent resonance energy transfer".

KEY WORDS:

Diagnostic method, Ribozyme, DNA-hairpin, SARS-CoV-2, FRET, HCR.

PAGES: 68
BIG AREA: Ciências Biológicas
AREA: Biologia Geral
SUMMARY:

The COVID-19 pandemic began in Wuhan province, located in China, and was caused by the SARS-CoV-2 coronavirus. Within a few months, the virus had already spread throughout the world, leading to a major global public health concern. To date, more than 770 million cases of infection and more than 6.9 million deaths have been recorded worldwide. These high infection numbers demonstrate the importance of diagnostic methods capable of identifying the presence of the virus in the first days of infection to guide public policies on containment measures to prevent its further spread. For this initial phase, the recommended method is RT-qPCR, currently considered the gold standard. However, this methodology presents some limitations such as high production cost, need for refrigeration, long execution time of the technique in addition to the need for specialized labor and equipment. Consequently, the need for refrigeration during transport and storage limits its wide distribution and use in various environments, especially those far from urban centers that do not have an adequate public health structure. Therefore, this study aims to develop a sensitive and easy-to-use diagnostic method, free from enzymatic activity, specialized equipment, labor and refrigeration. Relying on ribozymes for the recognition and cleavage of viral RNA and metastable DNA hairpin molecules with fluorophores at their ends, capable of performing hybridization chain reaction followed by fluorescence resonant energy transfer (FRET-HCR) for identification of the virus. The results indicated that two engineered ribozymes were able to identify and cleave the in vitro transcribed target of viral RNA and the DNA hairpin molecules were efficient in HCR reactions and the fluorophores in FRET reactions, with the best results showing fluorescence intensity 1.34 ± 0,11 times higher than the negative control, thus highlighting the potential of the proposed method. Therefore, this study served as a proof of concept for the joint use of ribozymes and DNA hairpin with fluorophores in FRET-HCR reactions to detect SARS-CoV-2.

COMMITTEE MEMBERS:
Presidente - 1207115 - ILDINETE SILVA PEREIRA
Interno - 2122537 - MARCELO DE MACEDO BRIGIDO
Externo à Instituição - MARCELO HENRIQUE SOLLER RAMADA - UCB
Externo ao Programa - 7405042 - MARCIO JOSE POCAS FONSECA - null