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Fecal Microbiota Transplantation: What’s New?
Article

Validation of Two Commercial Multiplex Real-Time PCR Assays for Detection of SARS-CoV-2 in Stool Donors for Fecal Microbiota Transplantation

1
Department of Surgical Sciences and Integrated Diagnostics (DISC), University of Genoa, 16132 Genoa, Italy
2
Microbiology and Virology Unit, Florence Careggi University Hospital, 50134 Florence, Italy
3
Department of Experimental and Clinical Medicine, University of Florence, 50134 Florence, Italy
4
Department of Translational Research and of New Technologies in Medicine and Surgery, University of Pisa, 56127 Pisa, Italy
5
Department of Laboratory Sciences and Infectious Diseases, Fondazione Policlinico Universitario A. Gemelli IRCCS, 00168 Rome, Italy
6
Department of Medical Biotechnologies, University of Siena, 53100 Siena, Italy
7
Microbiology, Fondazione Policlinico Universitario Agostino Gemelli IRCCS, Catholic University of Sacred Heart, 00168 Rome, Italy
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Paolo Calistri
Microorganisms 2022, 10(2), 284; https://doi.org/10.3390/microorganisms10020284
Received: 24 December 2021 / Revised: 21 January 2022 / Accepted: 23 January 2022 / Published: 26 January 2022
(This article belongs to the Special Issue Impact of Fecal Microbiota Transplantation on Human Health)
Recurrent infection by Clostridioides difficile has recently been treated by fecal microbiota transplantation (FMT). As viable SARS-CoV-2 was recovered from stool of asymptomatic individuals, the FMT procedure could be a potential risk of SARS-CoV-2 transmission, thus underlying the need to reliably detect SARS-CoV-2 in stool. Here, we performed a multicentric study to explore performances of two commercially available assays for detection of SARS-CoV-2 RNA in stool of potential FMT donors. In three hospitals, 180 stool samples were spiked with serial 10-fold dilutions of a SARS-CoV-2 inactivated lysate to evaluate the Seegene Allplex™ SARS-CoV-2 (SC2) and SARS-CoV-2/FluA/FluB/RSV (SC2FABR) Assays for the detection of viral RNA in stool of FMT donors. The results revealed that both assays detected down to 2 TCID50/mL with comparable limit of detection values, SC2 showing more consistent target positivity rate than SC2FABR. Beyond high amplification efficiency, correlation between CT values and log concentrations of inactivated viral lysates showed R2 values ranging from 0.88 to 0.90 and from 0.87 to 0.91 for the SC2 and SC2FABR assay, respectively. The present results demonstrate that both methods are highly reproducible, sensitive, and accurate for SARS-CoV-2 RNA detection in stool, suggesting a potential use in FMT-donor screening. View Full-Text
Keywords: FMT; RT-PCR; COVID-19; feces; donor screening FMT; RT-PCR; COVID-19; feces; donor screening
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MDPI and ACS Style

Di Pilato, V.; Morecchiato, F.; Rizzato, C.; Quaranta, G.; Fais, R.; Gandolfo, C.; Antonelli, A.; Cusi, M.G.; Pistello, M.; Rossolini, G.M.; Sanguinetti, M.; Lupetti, A.; Masucci, L. Validation of Two Commercial Multiplex Real-Time PCR Assays for Detection of SARS-CoV-2 in Stool Donors for Fecal Microbiota Transplantation. Microorganisms 2022, 10, 284. https://doi.org/10.3390/microorganisms10020284

AMA Style

Di Pilato V, Morecchiato F, Rizzato C, Quaranta G, Fais R, Gandolfo C, Antonelli A, Cusi MG, Pistello M, Rossolini GM, Sanguinetti M, Lupetti A, Masucci L. Validation of Two Commercial Multiplex Real-Time PCR Assays for Detection of SARS-CoV-2 in Stool Donors for Fecal Microbiota Transplantation. Microorganisms. 2022; 10(2):284. https://doi.org/10.3390/microorganisms10020284

Chicago/Turabian Style

Di Pilato, Vincenzo, Fabio Morecchiato, Cosmeri Rizzato, Gianluca Quaranta, Roberta Fais, Claudia Gandolfo, Alberto Antonelli, Maria G. Cusi, Mauro Pistello, Gian M. Rossolini, Maurizio Sanguinetti, Antonella Lupetti, and Luca Masucci. 2022. "Validation of Two Commercial Multiplex Real-Time PCR Assays for Detection of SARS-CoV-2 in Stool Donors for Fecal Microbiota Transplantation" Microorganisms 10, no. 2: 284. https://doi.org/10.3390/microorganisms10020284

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