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SARS-CoV2 detection, in different biological specimens, among people infected with this virus

Grant number: 20/05623-0
Support type:Regular Research Grants
Duration: February 01, 2021 - January 31, 2023
Field of knowledge:Health Sciences - Medicine - Medical Clinics
Principal researcher:Maria Cássia Jacintho Mendes Corrêa
Grantee:Maria Cássia Jacintho Mendes Corrêa
Home Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Assoc. researchers: Ana Luiza de Souza Bierrenbach ; Fabio Eudes Leal ; João Carlos da Silva Bizario ; Silvia Figueiredo Costa ; Tania Regina Tozetto Mendoza

Abstract

SARS-CoV-2 has been detected from a variety of upper and lower respiratory sources including throat, nasal/ nasopharyngeal, sputum, and bronchial fluid during the acute phase of the disease. However, little is known about the presence of this virus in other biological specimens and at different phases of the disease. Likewise, little is known about the association of the excretion of this agent in these secretions and the immune-humoral responses type of information could assist us in understanding the natural history of the disease and additionally could clarify aspects of interpersonal, hospital and community transmission of this virus. Objectives 1- To investigate the presence of SARS-CoV-2 in nasopharyngeal swab, saliva, urine, blood and feces, during the acute and convalescent phase of the infection, in immunocompetent and immunocompromised patients ; 2-To investigate the presence of SARS-CoV-2 in saliva or naso-pharyngeal swab of contacts of patients with Covid-19; 3- Correlate the presence of SARS-CoV 2 in these biological materials and the titers of neutralizing antibodies and IgG antibodies by ELISA; 4- To investigate the SARS-CoV-2 permanence and infectivity rate in these patients during the acute and convalescent phase of COVID-19Methods- This is an observational, prospective study, to be carried out on patients and asymptomatic contacts of these patients, seen at the Hospital das Clínicas da Faculdade de Medicina da USP, at the Cancer Institute of the State of São Paulo (ICESP) and in the municipality of São Caetano do Sul, diagnosed with SARS-CoV-2 infection.125 patients will be included, divided into 3 groups of patients: 1-Symptomatic patients without diagnosis of any previous immunodepression (50 patients); 2-Symptomatic patients diagnosed with solid tumors or candidates for Hematopoietic Stem Cell Transplantation under follow-up at ICESP (25 patients); 3-Asymptomatic contacts of these patients (any group-50 patients). Blood, urine, saliva, naso pharyngeal and anal swabs will be collected for 5 consecutive weeks, with intervals of one week (days 1, 7,14,21, 28) starting up to 72 hours from the time of diagnosis in all patients in groups 1 and 2. For patients in group 3, only saliva and naso pharynx swab will be collected on day 1. For the detection of anti-SARS CoV 2 antibodies, ELISA serological techniques and the neutralization assay will be used. For viral identification, molecular biology and viral culture techniques will be used. A quantitative duplex assay (internal control and specific RdRP-P2 gene) for SARS-CoV-2 will be performed by the real-time PCR assay, according to a protocol adapted from Charité, Berlin, with the primers and probes for the specific PCR aimed at amplification of the RdRP-P2 gene. Viral isolation will be performed in a security laboratory level 3 (NB-3), and the samples will be subjected to isolation in VeroE6 cells, known to be susceptible to Coronavirus. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MENDES CORREA, MARIA C.; LEAL, FABIO E.; VILLAS BOAS, LUCY S.; WITKIN, STEVEN S.; DE PAULA, ANDERSON; TOZETTO MENDONZA, TANIA R.; FERREIRA, NOELY E.; CURTY, GISLAINE; DE CARVALHO, PEDRO S.; BUSS, LEWIS F.; COSTA, SILVIA F.; DA CUNHA CARVALHO, FLAVIA M.; KAWAKAMI, JOYCE; TANIWAKI, NOEMI N.; PAIAO, HEUDER; DA SILVA BIZARIO, JOAO C.; DE JESUS, JAQUELINE G.; SABINO, ESTER C.; ROMANO, CAMILA M.; GREPAN, REGINA M. Z.; SESSO, ANTONIO. Prolonged presence of replication-competent SARS-CoV-2 in mildly symptomatic individuals: A report of two cases. Journal of Medical Virology, v. 93, n. 9, p. 5603-5607, SEP 2021. Web of Science Citations: 1.

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