Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung Samples

The Coronavirus Disease 19 (COVID-19) pandemic has caused an unexpected death toll worldwide. Even though several guidelines for the management of infectious corpses have been proposed, the limited number of post-mortem analyses during the pandemic has led to inaccuracies in the counting of COVID-19...

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Main Authors: Nicolò Musso, Luca Falzone, Stefano Stracquadanio, Dafne Bongiorno, Monica Salerno, Massimiliano Esposito, Francesco Sessa, Massimo Libra, Stefania Stefani, Cristoforo Pomara
Format: Article
Language:English
Published: MDPI AG 2021-06-01
Series:Diagnostics
Subjects:
Online Access:https://www.mdpi.com/2075-4418/11/7/1158
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author Nicolò Musso
Luca Falzone
Stefano Stracquadanio
Dafne Bongiorno
Monica Salerno
Massimiliano Esposito
Francesco Sessa
Massimo Libra
Stefania Stefani
Cristoforo Pomara
author_facet Nicolò Musso
Luca Falzone
Stefano Stracquadanio
Dafne Bongiorno
Monica Salerno
Massimiliano Esposito
Francesco Sessa
Massimo Libra
Stefania Stefani
Cristoforo Pomara
author_sort Nicolò Musso
collection DOAJ
description The Coronavirus Disease 19 (COVID-19) pandemic has caused an unexpected death toll worldwide. Even though several guidelines for the management of infectious corpses have been proposed, the limited number of post-mortem analyses during the pandemic has led to inaccuracies in the counting of COVID-19 deaths and contributed to a lack of important information about the pathophysiology of the SARS-CoV-2 infection. Due to the impossibility of carrying out autopsies on all corpses, the scientific community has raised the question of whether confirmatory analyses could be performed on exhumed bodies after a long period of burial to assess the presence of SARS-CoV-2 RNA. Post-mortem lung samples were collected from 16 patients who died from COVID-19 infection and were buried for a long period of time. A custom RNA extraction protocol was developed to enhance extraction of viral RNA from degraded samples and highly sensitive molecular methods, including RT-qPCR and droplet digital PCR (ddPCR), were used to detect the presence of SARS-CoV-2 RNA. The custom extraction protocol developed allowed us to extract total RNA effectively from all lung samples collected. SARS-CoV-2 viral RNA was effectively detected in all samples by both RT-qPCR and ddPCR, regardless of the length of burial. ddPCR results confirmed the persistence of the virus in this anatomical niche and revealed high viral loads in some lung samples, suggesting active infection at the time of death. To the best of our knowledge, this is the first study to demonstrate the persistence of SARS-CoV-2 viral RNA in the lung even after a long post-mortem interval (up to 78 days). The extraction protocol herein described, and the highly sensitive molecular analyses performed, could represent the standard procedures for SARS-CoV-2 detection in degraded lung specimens. Finally, the innovative results obtained encourage post-mortem confirmatory analyses even after a long post-mortem interval.
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spelling doaj.art-386ceb1abb7e40169310dca167f373912023-11-22T01:35:11ZengMDPI AGDiagnostics2075-44182021-06-01117115810.3390/diagnostics11071158Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung SamplesNicolò Musso0Luca Falzone1Stefano Stracquadanio2Dafne Bongiorno3Monica Salerno4Massimiliano Esposito5Francesco Sessa6Massimo Libra7Stefania Stefani8Cristoforo Pomara9Laboratory of Molecular and Resistant Antibiotic Medical Microbiology (MMAR), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, ItalyLaboratory of Experimental Oncology, Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, ItalyLaboratory of Molecular and Resistant Antibiotic Medical Microbiology (MMAR), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, ItalyLaboratory of Molecular and Resistant Antibiotic Medical Microbiology (MMAR), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, ItalyDepartment of Medical and Surgical Sciences and Advanced Technologies “G.F. Ingrassia”, Institute of Legal Medicine, University of Catania, 95123 Catania, ItalyDepartment of Medical and Surgical Sciences and Advanced Technologies “G.F. Ingrassia”, Institute of Legal Medicine, University of Catania, 95123 Catania, ItalyDepartment of Clinical and Experimental Medicine, University of Foggia, 71122 Foggia, ItalyLaboratory of Experimental Oncology, Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, ItalyLaboratory of Molecular and Resistant Antibiotic Medical Microbiology (MMAR), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, ItalyDepartment of Medical and Surgical Sciences and Advanced Technologies “G.F. Ingrassia”, Institute of Legal Medicine, University of Catania, 95123 Catania, ItalyThe Coronavirus Disease 19 (COVID-19) pandemic has caused an unexpected death toll worldwide. Even though several guidelines for the management of infectious corpses have been proposed, the limited number of post-mortem analyses during the pandemic has led to inaccuracies in the counting of COVID-19 deaths and contributed to a lack of important information about the pathophysiology of the SARS-CoV-2 infection. Due to the impossibility of carrying out autopsies on all corpses, the scientific community has raised the question of whether confirmatory analyses could be performed on exhumed bodies after a long period of burial to assess the presence of SARS-CoV-2 RNA. Post-mortem lung samples were collected from 16 patients who died from COVID-19 infection and were buried for a long period of time. A custom RNA extraction protocol was developed to enhance extraction of viral RNA from degraded samples and highly sensitive molecular methods, including RT-qPCR and droplet digital PCR (ddPCR), were used to detect the presence of SARS-CoV-2 RNA. The custom extraction protocol developed allowed us to extract total RNA effectively from all lung samples collected. SARS-CoV-2 viral RNA was effectively detected in all samples by both RT-qPCR and ddPCR, regardless of the length of burial. ddPCR results confirmed the persistence of the virus in this anatomical niche and revealed high viral loads in some lung samples, suggesting active infection at the time of death. To the best of our knowledge, this is the first study to demonstrate the persistence of SARS-CoV-2 viral RNA in the lung even after a long post-mortem interval (up to 78 days). The extraction protocol herein described, and the highly sensitive molecular analyses performed, could represent the standard procedures for SARS-CoV-2 detection in degraded lung specimens. Finally, the innovative results obtained encourage post-mortem confirmatory analyses even after a long post-mortem interval.https://www.mdpi.com/2075-4418/11/7/1158COVID-19autopsySARS-CoV-2post-mortem swabviral RNA
spellingShingle Nicolò Musso
Luca Falzone
Stefano Stracquadanio
Dafne Bongiorno
Monica Salerno
Massimiliano Esposito
Francesco Sessa
Massimo Libra
Stefania Stefani
Cristoforo Pomara
Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung Samples
Diagnostics
COVID-19
autopsy
SARS-CoV-2
post-mortem swab
viral RNA
title Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung Samples
title_full Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung Samples
title_fullStr Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung Samples
title_full_unstemmed Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung Samples
title_short Post-Mortem Detection of SARS-CoV-2 RNA in Long-Buried Lung Samples
title_sort post mortem detection of sars cov 2 rna in long buried lung samples
topic COVID-19
autopsy
SARS-CoV-2
post-mortem swab
viral RNA
url https://www.mdpi.com/2075-4418/11/7/1158
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