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Droplet digital PCR application for the detection of SARS-CoV-2 in air sample

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) may transmit through airborne route particularly when the aerosol particles remain in enclosed spaces with inadequate ventilation. There has been no standard recommended method of determining the virus in air due to limitations in pre-anal...

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Autores principales: Rashid, Siti Aishah, Nazakat, Raheel, Muhamad Robat, Rosnawati, Ismail, Rohaida, Suppiah, Jeyanthi, Rajendran, Kamesh, Raj Louis Masalamany, A. S. Santhana, Muhamad Hendri, Nur Afrina, Mohamad, Nadia, Khairul Hasni, Nurul Amalina, Suib, Fatin Amirah, Nik Hassan, Nik Muhamad Nizam, Pahrol, Muhammad Alfatih, Shaharudin, Rafiza
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641526/
https://www.ncbi.nlm.nih.gov/pubmed/37965510
http://dx.doi.org/10.3389/fpubh.2023.1208348
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author Rashid, Siti Aishah
Nazakat, Raheel
Muhamad Robat, Rosnawati
Ismail, Rohaida
Suppiah, Jeyanthi
Rajendran, Kamesh
Raj Louis Masalamany, A. S. Santhana
Muhamad Hendri, Nur Afrina
Mohamad, Nadia
Khairul Hasni, Nurul Amalina
Suib, Fatin Amirah
Nik Hassan, Nik Muhamad Nizam
Pahrol, Muhammad Alfatih
Shaharudin, Rafiza
author_facet Rashid, Siti Aishah
Nazakat, Raheel
Muhamad Robat, Rosnawati
Ismail, Rohaida
Suppiah, Jeyanthi
Rajendran, Kamesh
Raj Louis Masalamany, A. S. Santhana
Muhamad Hendri, Nur Afrina
Mohamad, Nadia
Khairul Hasni, Nurul Amalina
Suib, Fatin Amirah
Nik Hassan, Nik Muhamad Nizam
Pahrol, Muhammad Alfatih
Shaharudin, Rafiza
author_sort Rashid, Siti Aishah
collection PubMed
description Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) may transmit through airborne route particularly when the aerosol particles remain in enclosed spaces with inadequate ventilation. There has been no standard recommended method of determining the virus in air due to limitations in pre-analytical and technical aspects. Furthermore, the presence of low virus loads in air samples could result in false negatives. Our study aims to explore the feasibility of detecting SARS-CoV-2 ribonucleic acid (RNA) in air samples using droplet digital polymerase chain reaction (ddPCR). Active and passive air sampling was conducted between December 2021 and February 2022 with the presence of COVID-19 confirmed cases in two hospitals and a quarantine center in Klang Valley, Malaysia. SARS-CoV-2 RNA in air was detected and quantified using ddPCR and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The comparability of two different digital PCR platforms (QX200 and QIAcuity) to RT-PCR were also investigated. Additionally negative staining transmission electron microscopy was performed to visualize virus ultrastructure. Detection rates of SARS-CoV-2 in air samples using ddPCR were higher compared to RT-PCR, which were 15.2% (22/145) and 3.4% (5/145), respectively. The sensitivity and specificity of ddPCR was 100 and 87%, respectively. After excluding 17 negative samples (50%) by both QX200 and QIAcuity, 15% samples (5/34) were found to be positive both ddPCR and dPCR. There were 23.5% (8/34) samples that were detected positive by ddPCR but negative by dPCR. In contrast, there were 11.7% (4/34) samples that were detected positive by dPCR but negative by ddPCR. The SARS-CoV-2 detection method by ddPCR is precise and has a high sensitivity for viral RNA detection. It could provide advances in determining low viral titter in air samples to reduce false negative reports, which could complement detection by RT-PCR.
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spelling pubmed-106415262023-11-14 Droplet digital PCR application for the detection of SARS-CoV-2 in air sample Rashid, Siti Aishah Nazakat, Raheel Muhamad Robat, Rosnawati Ismail, Rohaida Suppiah, Jeyanthi Rajendran, Kamesh Raj Louis Masalamany, A. S. Santhana Muhamad Hendri, Nur Afrina Mohamad, Nadia Khairul Hasni, Nurul Amalina Suib, Fatin Amirah Nik Hassan, Nik Muhamad Nizam Pahrol, Muhammad Alfatih Shaharudin, Rafiza Front Public Health Public Health Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) may transmit through airborne route particularly when the aerosol particles remain in enclosed spaces with inadequate ventilation. There has been no standard recommended method of determining the virus in air due to limitations in pre-analytical and technical aspects. Furthermore, the presence of low virus loads in air samples could result in false negatives. Our study aims to explore the feasibility of detecting SARS-CoV-2 ribonucleic acid (RNA) in air samples using droplet digital polymerase chain reaction (ddPCR). Active and passive air sampling was conducted between December 2021 and February 2022 with the presence of COVID-19 confirmed cases in two hospitals and a quarantine center in Klang Valley, Malaysia. SARS-CoV-2 RNA in air was detected and quantified using ddPCR and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The comparability of two different digital PCR platforms (QX200 and QIAcuity) to RT-PCR were also investigated. Additionally negative staining transmission electron microscopy was performed to visualize virus ultrastructure. Detection rates of SARS-CoV-2 in air samples using ddPCR were higher compared to RT-PCR, which were 15.2% (22/145) and 3.4% (5/145), respectively. The sensitivity and specificity of ddPCR was 100 and 87%, respectively. After excluding 17 negative samples (50%) by both QX200 and QIAcuity, 15% samples (5/34) were found to be positive both ddPCR and dPCR. There were 23.5% (8/34) samples that were detected positive by ddPCR but negative by dPCR. In contrast, there were 11.7% (4/34) samples that were detected positive by dPCR but negative by ddPCR. The SARS-CoV-2 detection method by ddPCR is precise and has a high sensitivity for viral RNA detection. It could provide advances in determining low viral titter in air samples to reduce false negative reports, which could complement detection by RT-PCR. Frontiers Media S.A. 2023-10-27 /pmc/articles/PMC10641526/ /pubmed/37965510 http://dx.doi.org/10.3389/fpubh.2023.1208348 Text en Copyright © 2023 Rashid, Nazakat, Muhamad Robat, Ismail, Suppiah, Rajendran, Raj Louis Masalamany, Muhamad Hendri, Mohamad, Khairul Hasni, Suib, Nik Hassan, Pahrol and Shaharudin. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Public Health
Rashid, Siti Aishah
Nazakat, Raheel
Muhamad Robat, Rosnawati
Ismail, Rohaida
Suppiah, Jeyanthi
Rajendran, Kamesh
Raj Louis Masalamany, A. S. Santhana
Muhamad Hendri, Nur Afrina
Mohamad, Nadia
Khairul Hasni, Nurul Amalina
Suib, Fatin Amirah
Nik Hassan, Nik Muhamad Nizam
Pahrol, Muhammad Alfatih
Shaharudin, Rafiza
Droplet digital PCR application for the detection of SARS-CoV-2 in air sample
title Droplet digital PCR application for the detection of SARS-CoV-2 in air sample
title_full Droplet digital PCR application for the detection of SARS-CoV-2 in air sample
title_fullStr Droplet digital PCR application for the detection of SARS-CoV-2 in air sample
title_full_unstemmed Droplet digital PCR application for the detection of SARS-CoV-2 in air sample
title_short Droplet digital PCR application for the detection of SARS-CoV-2 in air sample
title_sort droplet digital pcr application for the detection of sars-cov-2 in air sample
topic Public Health
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10641526/
https://www.ncbi.nlm.nih.gov/pubmed/37965510
http://dx.doi.org/10.3389/fpubh.2023.1208348
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