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An optimised protocol for detection of SARS-CoV-2 in stool

An optimised protocol for detection of SARS-CoV-2 in stool

Authors :
Tianqi Li
Enriqueta Garcia-Gutierrez
Daniel A. Yara
Jacob Scadden
Jade Davies
Chloe Hutchins
Alp Aydin
Justin O’Grady
Arjan Narbad
Stefano Romano
Lizbeth Sayavedra
Source :
BMC Microbiology, Vol 21, Iss 1, Pp 1-8 (2021)
Publication Year :
2021
Publisher :
BMC, 2021.

Abstract

Abstract Background SARS-CoV-2 has been detected in stool samples of COVID-19 patients, with potential implications for faecal-oral transmission. Compared to nasopharyngeal swab samples, the complexity of the stool matrix poses a challenge in the detection of the virus that has not yet been solved. However, robust and reliable methods are needed to estimate the prevalence and persistence of SARS-CoV-2 in the gut and to ensure the safety of microbiome-based procedures such as faecal microbiota transplant (FMT). The aim of this study was to establish a sensitive and reliable method for detecting SARS-CoV-2 in stool samples. Results Stool samples from individuals free of SARS-CoV-2 were homogenised in saline buffer and spiked with a known titre of inactivated virus ranging from 50 to 750 viral particles per 100 mg stool. Viral particles were concentrated by ultrafiltration, RNA was extracted, and SARS-CoV-2 was detected via real-time reverse-transcription polymerase chain reaction (RT-qPCR) using the CDC primers and probes. The RNA extraction procedure we used allowed for the detection of SARS-CoV-2 via RT-qPCR in most of the stool samples tested. We could detect as few as 50 viral particles per 100 mg of stool. However, high variability was observed across samples at low viral titres. The primer set targeting the N1 region provided more reliable and precise results and for this primer set our method had a limit of detection of 1 viral particle per mg of stool. Conclusions Here we describe a sensitive method for detecting SARS-CoV-2 in stool samples. This method can be used to establish the persistence of SARS-CoV-2 in stool and ensure the safety of clinical practices such as FMT.

Details

Language :
English
ISSN :
14712180
Volume :
21
Issue :
1
Database :
Directory of Open Access Journals
Journal :
BMC Microbiology
Publication Type :
Academic Journal
Accession number :
edsdoj.68b9cb353b14be58a3ed5940cd45e4a
Document Type :
article
Full Text :
https://doi.org/10.1186/s12866-021-02297-w