key: cord-0931034-hawfavj1
authors: Fumian, Tulio M.; Malta, Fábio C.; dos Santos, Débora R. L.; Pauvolid‐Corrêa, Alex; Fialho, Alexandre M.; Leite, José P. G.; Miagostovich, Marize P.
title: SARS‐CoV‐2 RNA detection in stool samples from acute gastroenteritis cases, Brazil
date: 2021-01-22
journal: J Med Virol
DOI: 10.1002/jmv.26786
sha: be3f31e3f2e7e98c0207fd2e7806ea4181985147
doc_id: 931034
cord_uid: hawfavj1

We described the detection of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) in stool samples from patients presenting only acute gastroenteritis (AGE) symptoms. From January to July 2020, 121 AGE stool samples were screened by quantitative reverse‐transcription polymerase chain reaction. We detected SARS‐CoV‐2 in 27.5% of samples received during the epidemic period. No infectious viruses were observed in Vero E6 cells.

COVID-19 will show only moderate disease. However, a small fraction of patients will develop serious complications, progressing from severe pneumonia to acute respiratory distress syndrome, septic shock, and/or multiple organ failure. 5, 6 Whilst most of cases present as a classical respiratory disease, COVID-19 can include gastrointestinal, neurological, renal, and cardiovascular symptoms, as well as other complications. SARS-CoV-2 has been detected in organs other than the lungs, including the kidneys, liver, heart, brain, and blood. 7 Several studies have demonstrated the presence of the SARS-CoV-2 in stool samples from patients with or without diarrhea-associated symptoms. 8, 9 2 | MATERIAL AND METHODS Brazil's first COVID-19 case and death was respectively confirmed on February 25th and March 17th, 2020. From April onwards, it was observed a rapidly increase in the number of confirmed cases and deaths ( Figure 1 ). In the present study, we sought to investigated SARS-CoV-2 RNA in stool samples from acute gastroenteritis (AGE) cases before and during the COVID-19 epidemic in Brazil. For this purpose, we screened SARS-CoV-2 RNA in a total of 121 stool samples received between January and July 2020, previously tested negative for rotaviruses A (RVA) and noroviruses. Stool samples were retrospectively tested from the AGE biospecimen repository obtained from the Regional Rotavirus TaqMan®-based RT-qPCR assays, with primers and probe targeting the conserved NSP3 segment and ORF1/2 junction region, respectively, as previously described. 11, 12 Positive stool samples, stored at −80°C, were processed for viral isolation in cell culture inside a biosafety Level 3 facility. Briefly, stool suspensions were filtered (0.22 μm, Millex-GP Syringe Filter Unit, Millipore) and treated with 1% penicillin/streptomycin (P/S) and 0.5 μg/ml

Eagle's Medium supplemented with 5% fetal bovine serum, 1% penicillin/ streptomycin (P/S), 0.5 μg/ml Amphotericin B and 1% L-glutamine, were inoculated and incubated at 37°C with 5% CO 2 for 72 h. Following incubation of 3 days and up to the third passage, cytopathic effect (CPE) was evaluated under a microscope. Additionally, to check for SARS-CoV-2 replication, supernatant from the three passages were collected, RNA extracted and tested using RT-qPCR, as described above. | 2545

male patient showing diarrhea as the only onset symptom that REFERENCES 1. Coronavirus Resource Center

World Health Organization. Coronavirus disease 2019 (COVID-19) Dashboard

China Novel Coronavirus Investigating and Research Team. A novel coronavirus from patients with pneumonia in China

Enteric involvement of severe acute respiratory syndrome-associated coronavirus infection

Viral and host factors related to the clinical outcome of COVID-19

COVID-19: immunopathology and its implications for therapy

Multiorgan and renal tropism of SARS-CoV-2

Characteristics of pediatric SARS-CoV-2 infection and potential evidence for persistent fecal viral shedding

Evidence for gastrointestinal infection of SARS-CoV-2

US CDC real-time reverse transcription PCR panel for detection of severe acute respiratory syndrome coronavirus 2

One-step quantitative RT-PCR for the detection of rotavirus in acute gastroenteritis

Broadly reactive and highly sensitive assay for Norwalk-like viruses based on real-time quantitative reverse transcription-PCR

SARS-CoV-2 induced diarrhoea as onset symptom in patient with COVID-19

A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-toperson transmission: a study of a family cluster

Viral replication in the nasopharynx is associated with diarrhea in patients with severe acute respiratory syndrome

Middle East respiratory syndrome coronavirus: another zoonotic betacoronavirus causing SARS-like disease

Predicting infectious SARS-CoV-2 from diagnostic samples

Infectious SARS-CoV-2 in feces of patient with severe COVID-19

SARS-CoV-2 RNA detection in stool samples from acute gastroenteritis cases, Brazil

The authors declare that there are no conflict of interests. 

The data that support the findings of this study are available from the corresponding author upon reasonable request.

https://orcid.org/0000-0001-9654-0172