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Multi-modality detection of SARS-CoV-2 in faecal donor samples for transplantation and in asymptomatic emergency surgical admissions
Susan E Manzoor; Shafquat Zaman; Celina M Whalley; David Inglis; Andrew Bosworth; Michael Kidd; Sahida Shabir; Nabil Quraishi; Christopher A Green; Tariq H Iqbal; Andrew D Beggs.
Afiliación
  • Susan E Manzoor; University of Birmingham
  • Shafquat Zaman; University of Birmingham
  • Celina M Whalley; University of Birmingham
  • David Inglis; University of Birmingham
  • Andrew Bosworth; University of Birmingham
  • Michael Kidd; University of Birmingham
  • Sahida Shabir; University of Birmingham
  • Nabil Quraishi; University of Birmingham
  • Christopher A Green; University of Birmingham
  • Tariq H Iqbal; University of Birmingham
  • Andrew D Beggs; University of Birmingham
Preprint en En | PREPRINT-MEDRXIV | ID: ppmedrxiv-21250934
ABSTRACT
IntroductionFaecal transplantation is an evidence based treatment for Clostridiodes difficile. Patients infected with SARS-CoV-2 have been shown to shed the virus in stool for up to 33 days, well beyond the average clearance time for upper respiratory tract shedding. We carried out an analytical and clinical validation of reverse-transcriptase quantitative (RT-qPCR) as well as LAMP, LamPORE and droplet digital PCR in the detection of SARS-CoV-2 RNA in stool from donated samples for FMT, spiked samples and asymptomatic inpatients in an acute surgical unit. MethodsKilled SARS-CoV-2 viral lysate and extracted RNA was spiked into donor stool & FMT and a linear dilution series from 10-1 to 10-5 and tested via RT-qPCR, LAMP, LamPORE and ddPCR against SARS-CoV-2. Patients admitted to the critical care unit with symptomatic SARS-CoV-2 and sequential asymptomatic patients from acute presentation to an acute surgical unit were also tested. ResultsIn a linear dilution series, detection of the lowest dilution series was found to be 8 copies per microlitre of sample. Spiked lysate samples down to 10-2 dilution were detected in FMT samples using RTQPCR, LamPORE and ddPCR and down to 10-1 with LAMP. In symptomatic patients 5/12 had detectable SARS-CoV-2 in stool via RT-qPCR and 6/12 via LamPORE, and in 1/97 asymptomatic patients via RT-qPCR. ConclusionsRT-qPCR can be detected in FMT donor samples using RT-qPCR, LamPORE and ddPCR to low levels using validated pathways. As previously demonstrated, nearly half of symptomatic and less than one percent of asymptomatic patients had detectable SARS-CoV-2 in stool.
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Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-MEDRXIV Tipo de estudio: Prognostic_studies Idioma: En Año: 2021 Tipo del documento: Preprint
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-MEDRXIV Tipo de estudio: Prognostic_studies Idioma: En Año: 2021 Tipo del documento: Preprint