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Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits

BACKGROUND: Gastrointestinal symptoms are common in COVID-19 patients and SARS-CoV-2 RNA has been detected in the patients’ feces, which could lead to fecal–oral transmission. Therefore, fecal sample testing with real-time RT-PCR is highly recommended as a routine test for SARS-CoV-2 infection. Howe...

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Autores principales: Cao, Ranran, Bao, Lirong, Pan, Ming, Zhang, Cheng, Liao, Hongyu, Liu, Li, Li, Yan, Li, Mingyuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7893130/
https://www.ncbi.nlm.nih.gov/pubmed/33607939
http://dx.doi.org/10.1186/s12866-021-02118-0
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author Cao, Ranran
Bao, Lirong
Pan, Ming
Zhang, Cheng
Liao, Hongyu
Liu, Li
Li, Yan
Li, Mingyuan
author_facet Cao, Ranran
Bao, Lirong
Pan, Ming
Zhang, Cheng
Liao, Hongyu
Liu, Li
Li, Yan
Li, Mingyuan
author_sort Cao, Ranran
collection PubMed
description BACKGROUND: Gastrointestinal symptoms are common in COVID-19 patients and SARS-CoV-2 RNA has been detected in the patients’ feces, which could lead to fecal–oral transmission. Therefore, fecal sample testing with real-time RT-PCR is highly recommended as a routine test for SARS-CoV-2 infection. However, varying rates of detection in fecal sample have been reported. The aim of this study was to provide insights into the detection rates of SARS-CoV-2 in COVID-19 patients’ fecal sample by using four real-time RT-PCR kits and two pretreatment methods (inactive and non-inactive). RESULTS: The detection rate of Trizol pretreatment group was slightly higher than that of Phosphate Buffered Saline (PBS) groups, showing that pretreatment and inactivation by Trizol had no influence to SARS-CoV-2 nucleic acid test (NAT) results. 39.29% detection rate in fecal sample by DAAN was obtained, while Bio-germ was 40.48%, Sansure 34.52%, and GeneoDx 33.33%. The former three kits had no significant difference. The DAAN kit detection rates of ORF1ab and N gene were nearly equal and Ct value distribution was more scattered, while the Bio-germ kit distribution was more clustered. The positive rate of SARS-COV-2 in fecal samples correlated with the severity of the disease, specifically, severe cases were less likely to be identified than asymptomatic infection in the DAAN group (adjusted OR 0.05, 95%CI = 0.00 ~ 0.91). CONCLUSIONS: Trizol should be of choice as a valid and safe method for pretreatment of fecal samples of SARS-CoV-2. All real-time RT-PCR kits assessed in this study can be used for routine detection of SARS-CoV-2 in fecal samples. While DAAN, with high NAT positive rate, could be the best out of the 4 kits used in this study. SARS-CoV-2 positive rate in fecal sample was related to the severity of illness. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-021-02118-0.
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spelling pubmed-78931302021-02-19 Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits Cao, Ranran Bao, Lirong Pan, Ming Zhang, Cheng Liao, Hongyu Liu, Li Li, Yan Li, Mingyuan BMC Microbiol Research Article BACKGROUND: Gastrointestinal symptoms are common in COVID-19 patients and SARS-CoV-2 RNA has been detected in the patients’ feces, which could lead to fecal–oral transmission. Therefore, fecal sample testing with real-time RT-PCR is highly recommended as a routine test for SARS-CoV-2 infection. However, varying rates of detection in fecal sample have been reported. The aim of this study was to provide insights into the detection rates of SARS-CoV-2 in COVID-19 patients’ fecal sample by using four real-time RT-PCR kits and two pretreatment methods (inactive and non-inactive). RESULTS: The detection rate of Trizol pretreatment group was slightly higher than that of Phosphate Buffered Saline (PBS) groups, showing that pretreatment and inactivation by Trizol had no influence to SARS-CoV-2 nucleic acid test (NAT) results. 39.29% detection rate in fecal sample by DAAN was obtained, while Bio-germ was 40.48%, Sansure 34.52%, and GeneoDx 33.33%. The former three kits had no significant difference. The DAAN kit detection rates of ORF1ab and N gene were nearly equal and Ct value distribution was more scattered, while the Bio-germ kit distribution was more clustered. The positive rate of SARS-COV-2 in fecal samples correlated with the severity of the disease, specifically, severe cases were less likely to be identified than asymptomatic infection in the DAAN group (adjusted OR 0.05, 95%CI = 0.00 ~ 0.91). CONCLUSIONS: Trizol should be of choice as a valid and safe method for pretreatment of fecal samples of SARS-CoV-2. All real-time RT-PCR kits assessed in this study can be used for routine detection of SARS-CoV-2 in fecal samples. While DAAN, with high NAT positive rate, could be the best out of the 4 kits used in this study. SARS-CoV-2 positive rate in fecal sample was related to the severity of illness. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-021-02118-0. BioMed Central 2021-02-19 /pmc/articles/PMC7893130/ /pubmed/33607939 http://dx.doi.org/10.1186/s12866-021-02118-0 Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Cao, Ranran
Bao, Lirong
Pan, Ming
Zhang, Cheng
Liao, Hongyu
Liu, Li
Li, Yan
Li, Mingyuan
Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits
title Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits
title_full Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits
title_fullStr Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits
title_full_unstemmed Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits
title_short Detection of SARS-CoV-2 in fecal samples with different pretreatment methods and PCR kits
title_sort detection of sars-cov-2 in fecal samples with different pretreatment methods and pcr kits
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7893130/
https://www.ncbi.nlm.nih.gov/pubmed/33607939
http://dx.doi.org/10.1186/s12866-021-02118-0
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