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An optimised protocol for detection of SARS-CoV-2 in stool.
Li, Tianqi; Garcia-Gutierrez, Enriqueta; Yara, Daniel A; Scadden, Jacob; Davies, Jade; Hutchins, Chloe; Aydin, Alp; O'Grady, Justin; Narbad, Arjan; Romano, Stefano; Sayavedra, Lizbeth.
  • Li T; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Garcia-Gutierrez E; State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, China.
  • Yara DA; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Scadden J; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Davies J; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Hutchins C; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Aydin A; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • O'Grady J; Microbes in the Food Chain, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Narbad A; Microbes in the Food Chain, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Romano S; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK.
  • Sayavedra L; Gut Health and Microbes, Quadram Institute Bioscience, Norwich Research Park, Norwich, UK. Stefano.Romano@quadram.ac.uk.
BMC Microbiol ; 21(1): 242, 2021 09 06.
Article in English | MEDLINE | ID: covidwho-1398839
Preprint
This scientific journal article is probably based on a previously available preprint. It has been identified through a machine matching algorithm, human confirmation is still pending.
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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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Feces / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Observational study / Prognostic study Topics: Long Covid Limits: Humans Language: English Journal: BMC Microbiol Journal subject: Microbiology Year: 2021 Document Type: Article Affiliation country: S12866-021-02297-w

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Feces / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Observational study / Prognostic study Topics: Long Covid Limits: Humans Language: English Journal: BMC Microbiol Journal subject: Microbiology Year: 2021 Document Type: Article Affiliation country: S12866-021-02297-w