Dados do Trabalho

Título

METHODOLOGY FOR INVESTIGATION OF RBD DOMAIN OF GENE ENCODING SPIKE PROTEIN OF SARS-COV-2 IN THE UFABC COMMUNITY

Introdução

The SARS-CoV-2, a virus from the Coronaviridae family, emerged in Wuhan, China, in 2019 and is responsible for the COVID-19 pandemic. This virus presents a range of clinical manifestations, from asymptomatic infections to severe acute respiratory syndrome (SARS), affecting millions of people and causing numerous deaths. The genome of SARS-CoV-2 is composed of positive-sense RNA, protected by the nucleocapsid protein (N). The other structural proteins of the virus include the envelope (E) protein, membrane (M) protein, and the Spike protein (S). The Spike protein gives the virus its characteristic crown-like appearance and plays a crucial role in infection by binding to the angiotensin-converting enzyme 2 (ACE2) receptor to enter human cells. The S protein is the primary target of the host's immune response and the main component of available COVID-19 vaccines. The rapid evolution of RNA viruses contributes to regional specificities. The receptor-binding domain (RBD) of the S protein is crucial for receptor binding, and its modifications may affect the virus's ability to evade the immune response and vaccines.

Objetivo (s)

This study aimed to develop a method and protocol to facilitate the investigation of the genetic variability of the RBD region of the SARS-CoV-2 S protein in virus-positive samples collected during community monitoring at UFABC from May 2021 to December 2022. 

Material e Métodos

A total of 51,000 saliva samples were self-collected using cotton swabs by UFABC community members, with 1,729 testing positive for SARS-CoV-2. From these, a 404 bp fragment corresponding to the hypervariable region 1 (HV1) of the human mitochondrial gene was amplified by PCR to ensure sample quality. Subsequently, a 492 bp fragment of the S protein gene was amplified in 747 samples using PCR and Nested-PCR. Primers were designed and synthesized to amplify an 800 bp fragment of the RBD domain of the S protein from all 747 samples.

Resultados e Conclusão

The detection of SARS-CoV-2 and the characterization of its variants are crucial for guiding public health strategies and establishing sentinel communities for monitoring specific regions. This methodology, which detects the RBD domain, allows for the identification and monitoring of various virus strains and will serve as the basis for large-scale investigations into the genetic variability of the RBD domain of the SARS-CoV-2 S protein. 

Palavras Chave

SARS-CoV-2; COVID-19 pandemic; Receptor da Enzima Conversora de Angiotensina; Domínio de ligação ao receptor; leishmaniose