Viable SARS-CoV-2 in various specimens from COVID-19 patients

Objectives: The aim was to determine whether various clinical specimens obtained from COVID-19 pa-tients contain the infectious virus. Methods: To demonstrate whether various clinical specimens contain the viable virus, we collected naso/ oropharyngeal swabs and saliva, urine and stool samples from five COVID-19 patients and performed a quantitative polymerase chain reaction (qPCR) to assess viral load. Specimens positive with qPCR were subjected to virus isolation in Vero cells. We also used urine and stool samples to intranasally inoculate ferrets and evaluated the virus titres in nasal washes on 2, 4, 6 and 8 days post infection. Results: SARS-CoV-2 RNA was detected in all naso/oropharyngeal swabs and saliva, urine and stool samples collected between days 8 and 30 of the clinical course. Notably, viral loads in urine, saliva and stool samples were almost equal to or higher than those in naso/oropharyngeal swabs (urine 1.08 +/- 0.16-2.09 +/- 0.85 log(10) copies/mL, saliva 1.07 +/- 0.34-1.65 +/- 0.46 log(10) copies/mL, stool 1.17 +/- 0.32 log(10) copies/mL, naso/oropharyngeal swabs 1.18 +/- 0.12-1.34 +/- 0.30 log(10) copies/mL). Further, viable SARSCoV-2 was isolated from naso/oropharyngeal swabs and saliva of COVID-19 patients, as well as nasal washes of ferrets inoculated with patient urine or stool. Discussion: Viable SARS-CoV-2 was demonstrated in saliva, urine and stool samples from COVID-19 patients up to days 11-15 of the clinical course. This result suggests that viable SARS-CoV-2 can be secreted in various clinical samples and respiratory specimens. (c) 2020 The Author(s). Published by Elsevier Ltd on behalf of European Society of Clinical Microbiology and Infectious Diseases. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/).