Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay.
Identifieur interne : 000024 ( Main/Corpus ); précédent : 000023; suivant : 000025Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay.
Auteurs : Bernhard Schermer ; Francesca Fabretti ; Maximilian Damagnez ; Veronica Di Cristanziano ; Eva Heger ; Sita Arjune ; Nathan A. Tanner ; Thomas Imhof ; Manuel Koch ; Alim Ladha ; Julia Joung ; Jonathan S. Gootenberg ; Omar O. Abudayyeh ; Volker Burst ; Feng Zhang ; Florian Klein ; Thomas Benzing ; Roman-Ulrich MüllerSource :
- PloS one [ 1932-6203 ] ; 2020.
English descriptors
- KwdEn :
- Betacoronavirus (genetics), Betacoronavirus (isolation & purification), Clinical Laboratory Techniques (MeSH), Coronavirus Infections (diagnosis), Coronavirus Infections (virology), Humans (MeSH), Nasopharynx (virology), Nucleic Acid Amplification Techniques (methods), Pandemics (MeSH), Pneumonia, Viral (diagnosis), Pneumonia, Viral (virology), RNA, Viral (genetics), RNA, Viral (metabolism), Real-Time Polymerase Chain Reaction (MeSH), Sensitivity and Specificity (MeSH).
- MESH :
- chemical , genetics : RNA, Viral.
- diagnosis : Coronavirus Infections, Pneumonia, Viral.
- genetics : Betacoronavirus.
- isolation & purification : Betacoronavirus.
- chemical , metabolism : RNA, Viral.
- methods : Nucleic Acid Amplification Techniques.
- virology : Coronavirus Infections, Nasopharynx, Pneumonia, Viral.
- Clinical Laboratory Techniques, Humans, Pandemics, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity.
Abstract
BACKGROUND
Rapid and extensive testing of large parts of the population and specific subgroups is crucial for proper management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections and decision-making in times of a pandemic outbreak. However, point-of-care (POC) testing in places such as emergency units, outpatient clinics, airport security points or the entrance of any public building is a major challenge. The need for thermal cycling and nucleic acid isolation hampers the use of standard PCR-based methods for this purpose.
METHODS
To avoid these obstacles, we tested PCR-independent methods for the detection of SARS-CoV-2 RNA from primary material (nasopharyngeal swabs) including reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific high-sensitivity enzymatic reporter unlocking (SHERLOCK).
RESULTS
Whilst specificity of standard RT-LAMP assays appears to be satisfactory, sensitivity does not reach the current gold-standard quantitative real-time polymerase chain reaction (qPCR) assays yet. We describe a novel multiplexed RT-LAMP approach and validate its sensitivity on primary samples. This approach allows for fast and reliable identification of infected individuals. Primer optimization and multiplexing helps to increase sensitivity significantly. In addition, we directly compare and combine our novel RT-LAMP assays with SHERLOCK.
CONCLUSION
In summary, this approach reveals one-step multiplexed RT-LAMP assays as a prime-option for the development of easy and cheap POC test kits.
DOI: 10.1371/journal.pone.0238612
PubMed: 33137122
PubMed Central: PMC7605681
Links to Exploration step
pubmed:33137122Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay.</title><author><name sortKey="Schermer, Bernhard" sort="Schermer, Bernhard" uniqKey="Schermer B" first="Bernhard" last="Schermer">Bernhard Schermer</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Fabretti, Francesca" sort="Fabretti, Francesca" uniqKey="Fabretti F" first="Francesca" last="Fabretti">Francesca Fabretti</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Damagnez, Maximilian" sort="Damagnez, Maximilian" uniqKey="Damagnez M" first="Maximilian" last="Damagnez">Maximilian Damagnez</name><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Di Cristanziano, Veronica" sort="Di Cristanziano, Veronica" uniqKey="Di Cristanziano V" first="Veronica" last="Di Cristanziano">Veronica Di Cristanziano</name><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Heger, Eva" sort="Heger, Eva" uniqKey="Heger E" first="Eva" last="Heger">Eva Heger</name><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Arjune, Sita" sort="Arjune, Sita" uniqKey="Arjune S" first="Sita" last="Arjune">Sita Arjune</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Tanner, Nathan A" sort="Tanner, Nathan A" uniqKey="Tanner N" first="Nathan A" last="Tanner">Nathan A. Tanner</name><affiliation><nlm:affiliation>New England Biolabs, Ipswich, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Imhof, Thomas" sort="Imhof, Thomas" uniqKey="Imhof T" first="Thomas" last="Imhof">Thomas Imhof</name><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Koch, Manuel" sort="Koch, Manuel" uniqKey="Koch M" first="Manuel" last="Koch">Manuel Koch</name><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Ladha, Alim" sort="Ladha, Alim" uniqKey="Ladha A" first="Alim" last="Ladha">Alim Ladha</name><affiliation><nlm:affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Joung, Julia" sort="Joung, Julia" uniqKey="Joung J" first="Julia" last="Joung">Julia Joung</name><affiliation><nlm:affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Gootenberg, Jonathan S" sort="Gootenberg, Jonathan S" uniqKey="Gootenberg J" first="Jonathan S" last="Gootenberg">Jonathan S. Gootenberg</name><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Abudayyeh, Omar O" sort="Abudayyeh, Omar O" uniqKey="Abudayyeh O" first="Omar O" last="Abudayyeh">Omar O. Abudayyeh</name><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Burst, Volker" sort="Burst, Volker" uniqKey="Burst V" first="Volker" last="Burst">Volker Burst</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Emergency Department, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Zhang, Feng" sort="Zhang, Feng" uniqKey="Zhang F" first="Feng" last="Zhang">Feng Zhang</name><affiliation><nlm:affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Klein, Florian" sort="Klein, Florian" uniqKey="Klein F" first="Florian" last="Klein">Florian Klein</name><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>German Center for Infection Research, Partner Site Bonn-Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Benzing, Thomas" sort="Benzing, Thomas" uniqKey="Benzing T" first="Thomas" last="Benzing">Thomas Benzing</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Muller, Roman Ulrich" sort="Muller, Roman Ulrich" uniqKey="Muller R" first="Roman-Ulrich" last="Müller">Roman-Ulrich Müller</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2020">2020</date><idno type="RBID">pubmed:33137122</idno><idno type="pmid">33137122</idno><idno type="doi">10.1371/journal.pone.0238612</idno><idno type="pmc">PMC7605681</idno><idno type="wicri:Area/Main/Corpus">000024</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000024</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay.</title><author><name sortKey="Schermer, Bernhard" sort="Schermer, Bernhard" uniqKey="Schermer B" first="Bernhard" last="Schermer">Bernhard Schermer</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Fabretti, Francesca" sort="Fabretti, Francesca" uniqKey="Fabretti F" first="Francesca" last="Fabretti">Francesca Fabretti</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Damagnez, Maximilian" sort="Damagnez, Maximilian" uniqKey="Damagnez M" first="Maximilian" last="Damagnez">Maximilian Damagnez</name><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Di Cristanziano, Veronica" sort="Di Cristanziano, Veronica" uniqKey="Di Cristanziano V" first="Veronica" last="Di Cristanziano">Veronica Di Cristanziano</name><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Heger, Eva" sort="Heger, Eva" uniqKey="Heger E" first="Eva" last="Heger">Eva Heger</name><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Arjune, Sita" sort="Arjune, Sita" uniqKey="Arjune S" first="Sita" last="Arjune">Sita Arjune</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Tanner, Nathan A" sort="Tanner, Nathan A" uniqKey="Tanner N" first="Nathan A" last="Tanner">Nathan A. Tanner</name><affiliation><nlm:affiliation>New England Biolabs, Ipswich, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Imhof, Thomas" sort="Imhof, Thomas" uniqKey="Imhof T" first="Thomas" last="Imhof">Thomas Imhof</name><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Koch, Manuel" sort="Koch, Manuel" uniqKey="Koch M" first="Manuel" last="Koch">Manuel Koch</name><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Ladha, Alim" sort="Ladha, Alim" uniqKey="Ladha A" first="Alim" last="Ladha">Alim Ladha</name><affiliation><nlm:affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Joung, Julia" sort="Joung, Julia" uniqKey="Joung J" first="Julia" last="Joung">Julia Joung</name><affiliation><nlm:affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Gootenberg, Jonathan S" sort="Gootenberg, Jonathan S" uniqKey="Gootenberg J" first="Jonathan S" last="Gootenberg">Jonathan S. Gootenberg</name><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Abudayyeh, Omar O" sort="Abudayyeh, Omar O" uniqKey="Abudayyeh O" first="Omar O" last="Abudayyeh">Omar O. Abudayyeh</name><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Burst, Volker" sort="Burst, Volker" uniqKey="Burst V" first="Volker" last="Burst">Volker Burst</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Emergency Department, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Zhang, Feng" sort="Zhang, Feng" uniqKey="Zhang F" first="Feng" last="Zhang">Feng Zhang</name><affiliation><nlm:affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</nlm:affiliation></affiliation></author><author><name sortKey="Klein, Florian" sort="Klein, Florian" uniqKey="Klein F" first="Florian" last="Klein">Florian Klein</name><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>German Center for Infection Research, Partner Site Bonn-Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Benzing, Thomas" sort="Benzing, Thomas" uniqKey="Benzing T" first="Thomas" last="Benzing">Thomas Benzing</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author><author><name sortKey="Muller, Roman Ulrich" sort="Muller, Roman Ulrich" uniqKey="Muller R" first="Roman-Ulrich" last="Müller">Roman-Ulrich Müller</name><affiliation><nlm:affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</nlm:affiliation></affiliation><affiliation><nlm:affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</nlm:affiliation></affiliation></author></analytic><series><title level="j">PloS one</title><idno type="eISSN">1932-6203</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Betacoronavirus (genetics)</term><term>Betacoronavirus (isolation & purification)</term><term>Clinical Laboratory Techniques (MeSH)</term><term>Coronavirus Infections (diagnosis)</term><term>Coronavirus Infections (virology)</term><term>Humans (MeSH)</term><term>Nasopharynx (virology)</term><term>Nucleic Acid Amplification Techniques (methods)</term><term>Pandemics (MeSH)</term><term>Pneumonia, Viral (diagnosis)</term><term>Pneumonia, Viral (virology)</term><term>RNA, Viral (genetics)</term><term>RNA, Viral (metabolism)</term><term>Real-Time Polymerase Chain Reaction (MeSH)</term><term>Sensitivity and Specificity (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Coronavirus Infections</term><term>Pneumonia, Viral</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Betacoronavirus</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Betacoronavirus</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Nucleic Acid Amplification Techniques</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Coronavirus Infections</term><term>Nasopharynx</term><term>Pneumonia, Viral</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Clinical Laboratory Techniques</term><term>Humans</term><term>Pandemics</term><term>Real-Time Polymerase Chain Reaction</term><term>Sensitivity and Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b></p><p>Rapid and extensive testing of large parts of the population and specific subgroups is crucial for proper management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections and decision-making in times of a pandemic outbreak. However, point-of-care (POC) testing in places such as emergency units, outpatient clinics, airport security points or the entrance of any public building is a major challenge. The need for thermal cycling and nucleic acid isolation hampers the use of standard PCR-based methods for this purpose.</p></div><div type="abstract" xml:lang="en"><p><b>METHODS</b></p><p>To avoid these obstacles, we tested PCR-independent methods for the detection of SARS-CoV-2 RNA from primary material (nasopharyngeal swabs) including reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific high-sensitivity enzymatic reporter unlocking (SHERLOCK).</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>Whilst specificity of standard RT-LAMP assays appears to be satisfactory, sensitivity does not reach the current gold-standard quantitative real-time polymerase chain reaction (qPCR) assays yet. We describe a novel multiplexed RT-LAMP approach and validate its sensitivity on primary samples. This approach allows for fast and reliable identification of infected individuals. Primer optimization and multiplexing helps to increase sensitivity significantly. In addition, we directly compare and combine our novel RT-LAMP assays with SHERLOCK.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSION</b></p><p>In summary, this approach reveals one-step multiplexed RT-LAMP assays as a prime-option for the development of easy and cheap POC test kits.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">33137122</PMID><DateCompleted><Year>2020</Year><Month>11</Month><Day>18</Day></DateCompleted><DateRevised><Year>2020</Year><Month>11</Month><Day>18</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Electronic">1932-6203</ISSN><JournalIssue CitedMedium="Internet"><Volume>15</Volume><Issue>11</Issue><PubDate><Year>2020</Year></PubDate></JournalIssue><Title>PloS one</Title><ISOAbbreviation>PLoS One</ISOAbbreviation></Journal><ArticleTitle>Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay.</ArticleTitle><Pagination><MedlinePgn>e0238612</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1371/journal.pone.0238612</ELocationID><Abstract><AbstractText Label="BACKGROUND">Rapid and extensive testing of large parts of the population and specific subgroups is crucial for proper management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections and decision-making in times of a pandemic outbreak. However, point-of-care (POC) testing in places such as emergency units, outpatient clinics, airport security points or the entrance of any public building is a major challenge. The need for thermal cycling and nucleic acid isolation hampers the use of standard PCR-based methods for this purpose.</AbstractText><AbstractText Label="METHODS">To avoid these obstacles, we tested PCR-independent methods for the detection of SARS-CoV-2 RNA from primary material (nasopharyngeal swabs) including reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific high-sensitivity enzymatic reporter unlocking (SHERLOCK).</AbstractText><AbstractText Label="RESULTS">Whilst specificity of standard RT-LAMP assays appears to be satisfactory, sensitivity does not reach the current gold-standard quantitative real-time polymerase chain reaction (qPCR) assays yet. We describe a novel multiplexed RT-LAMP approach and validate its sensitivity on primary samples. This approach allows for fast and reliable identification of infected individuals. Primer optimization and multiplexing helps to increase sensitivity significantly. In addition, we directly compare and combine our novel RT-LAMP assays with SHERLOCK.</AbstractText><AbstractText Label="CONCLUSION">In summary, this approach reveals one-step multiplexed RT-LAMP assays as a prime-option for the development of easy and cheap POC test kits.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Schermer</LastName><ForeName>Bernhard</ForeName><Initials>B</Initials><Identifier Source="ORCID">0000-0002-5194-9000</Identifier><AffiliationInfo><Affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fabretti</LastName><ForeName>Francesca</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Damagnez</LastName><ForeName>Maximilian</ForeName><Initials>M</Initials><Identifier Source="ORCID">0000-0002-2745-7719</Identifier><AffiliationInfo><Affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Di Cristanziano</LastName><ForeName>Veronica</ForeName><Initials>V</Initials><AffiliationInfo><Affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Heger</LastName><ForeName>Eva</ForeName><Initials>E</Initials><AffiliationInfo><Affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Arjune</LastName><ForeName>Sita</ForeName><Initials>S</Initials><Identifier Source="ORCID">0000-0002-6121-4614</Identifier><AffiliationInfo><Affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tanner</LastName><ForeName>Nathan A</ForeName><Initials>NA</Initials><AffiliationInfo><Affiliation>New England Biolabs, Ipswich, MA, United States of America.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Imhof</LastName><ForeName>Thomas</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Koch</LastName><ForeName>Manuel</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Institute for Dental Research and Oral Musculoskeletal Biology, Center for Biochemistry, University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ladha</LastName><ForeName>Alim</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Joung</LastName><ForeName>Julia</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Gootenberg</LastName><ForeName>Jonathan S</ForeName><Initials>JS</Initials><AffiliationInfo><Affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Abudayyeh</LastName><ForeName>Omar O</ForeName><Initials>OO</Initials><AffiliationInfo><Affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Burst</LastName><ForeName>Volker</ForeName><Initials>V</Initials><AffiliationInfo><Affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Emergency Department, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Feng</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Howard Hughes Medical Institute, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>McGovern Institute for Brain Research at Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Massachusetts Consortium for Pathogen Readiness, Boston, MA, United States of America.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, United States of America.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Klein</LastName><ForeName>Florian</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Institute of Virology, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>German Center for Infection Research, Partner Site Bonn-Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Benzing</LastName><ForeName>Thomas</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Müller</LastName><ForeName>Roman-Ulrich</ForeName><Initials>RU</Initials><AffiliationInfo><Affiliation>Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Center for Molecular Medicine Cologne, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>11</Month><Day>02</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>PLoS One</MedlineTA><NlmUniqueID>101285081</NlmUniqueID><ISSNLinking>1932-6203</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012367">RNA, Viral</NameOfSubstance></Chemical></ChemicalList><SupplMeshList><SupplMeshName Type="Disease" UI="C000657245">COVID-19</SupplMeshName><SupplMeshName Type="Protocol" UI="C000657964">COVID-19 diagnostic testing</SupplMeshName><SupplMeshName Type="Organism" UI="C000656484">severe acute respiratory syndrome coronavirus 2</SupplMeshName></SupplMeshList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000073640" MajorTopicYN="N">Betacoronavirus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D019411" MajorTopicYN="N">Clinical Laboratory Techniques</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018352" MajorTopicYN="N">Coronavirus Infections</DescriptorName><QualifierName UI="Q000175" MajorTopicYN="Y">diagnosis</QualifierName><QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009305" MajorTopicYN="N">Nasopharynx</DescriptorName><QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D021141" MajorTopicYN="N">Nucleic Acid Amplification Techniques</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D058873" MajorTopicYN="N">Pandemics</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011024" MajorTopicYN="N">Pneumonia, Viral</DescriptorName><QualifierName UI="Q000175" MajorTopicYN="Y">diagnosis</QualifierName><QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012367" MajorTopicYN="N">RNA, Viral</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D060888" MajorTopicYN="N">Real-Time Polymerase Chain Reaction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012680" MajorTopicYN="N">Sensitivity and Specificity</DescriptorName></MeshHeading></MeshHeadingList><CoiStatement>F.Z., O.O.A., J.S.G., J.J., and A.L. are inventors on patent applications related to SHERLOCK technology filed by the Broad Institute, with the specific aim of ensuring this technology can be made freely, widely, and rapidly available for research and deployment (US provisional application no. 62/975,743, "CRISPR Effector System Based Coronavirus Diagnostics"). This does not alter our adherence to PLOS ONE`s policies on sharing data and materials. O.O.A., J.S.G., and F.Z. are co-founders, scientific advisors, and hold equity interests in Sherlock Biosciences, Inc., and Pine Trees Health, Inc. F.Z. is also a co-founder of Editas Medicine, Beam Therapeutics, Pairwise Plants, and Arbor Biotechnologies. NAT is employed by New England Biolabs, Inc, manufacturer of the LAMP reagents and some additional enzymes described in this manuscript. All of the aforementioned does not alter our adherence to PLOS ONE`s policies on sharing data and materials. The remaining authors declare no competing financial interests.</CoiStatement></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2020</Year><Month>08</Month><Day>25</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2020</Year><Month>10</Month><Day>14</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>11</Month><Day>2</Day><Hour>17</Hour><Minute>13</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>11</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>11</Month><Day>20</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">33137122</ArticleId><ArticleId IdType="doi">10.1371/journal.pone.0238612</ArticleId><ArticleId IdType="pii">PONE-D-20-26630</ArticleId><ArticleId IdType="pmc">PMC7605681</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>Euro Surveill. 2020 Apr;25(14):</Citation><ArticleIdList><ArticleId IdType="pubmed">32290902</ArticleId></ArticleIdList></Reference><Reference><Citation>Euro Surveill. 2020 Jan;25(3):</Citation><ArticleIdList><ArticleId IdType="pubmed">31992387</ArticleId></ArticleIdList></Reference><Reference><Citation>N Engl J Med. 2020 Sep 24;383(13):1283-1286</Citation><ArticleIdList><ArticleId IdType="pubmed">32857487</ArticleId></ArticleIdList></Reference><Reference><Citation>Nat Biotechnol. 2020 Jul;38(7):870-874</Citation><ArticleIdList><ArticleId IdType="pubmed">32300245</ArticleId></ArticleIdList></Reference><Reference><Citation>Microb Biotechnol. 2020 Jul;13(4):950-961</Citation><ArticleIdList><ArticleId IdType="pubmed">32333644</ArticleId></ArticleIdList></Reference><Reference><Citation>Virol Sin. 2020 Jun;35(3):344-347</Citation><ArticleIdList><ArticleId IdType="pubmed">32239445</ArticleId></ArticleIdList></Reference><Reference><Citation>Nat Med. 2020 May;26(5):672-675</Citation><ArticleIdList><ArticleId IdType="pubmed">32296168</ArticleId></ArticleIdList></Reference><Reference><Citation>Lancet Infect Dis. 2020 May;20(5):565-574</Citation><ArticleIdList><ArticleId IdType="pubmed">32213337</ArticleId></ArticleIdList></Reference><Reference><Citation>Eur J Clin Microbiol Infect Dis. 2020 Jun;39(6):1059-1061</Citation><ArticleIdList><ArticleId IdType="pubmed">32342252</ArticleId></ArticleIdList></Reference><Reference><Citation>Proc Natl Acad Sci U S A. 2020 Sep 15;117(37):22727-22735</Citation><ArticleIdList><ArticleId IdType="pubmed">32868442</ArticleId></ArticleIdList></Reference><Reference><Citation>Biotechniques. 2020 Sep;69(3):178-185</Citation><ArticleIdList><ArticleId IdType="pubmed">32635743</ArticleId></ArticleIdList></Reference><Reference><Citation>Int J Mol Sci. 2020 Apr 18;21(8):</Citation><ArticleIdList><ArticleId IdType="pubmed">32325642</ArticleId></ArticleIdList></Reference><Reference><Citation>J Mol Diagn. 2020 Jun;22(6):729-735</Citation><ArticleIdList><ArticleId IdType="pubmed">32276051</ArticleId></ArticleIdList></Reference><Reference><Citation>Nat Protoc. 2019 Oct;14(10):2986-3012</Citation><ArticleIdList><ArticleId IdType="pubmed">31548639</ArticleId></ArticleIdList></Reference><Reference><Citation>Emerg Microbes Infect. 2020 Dec;9(1):998-1007</Citation><ArticleIdList><ArticleId IdType="pubmed">32306853</ArticleId></ArticleIdList></Reference><Reference><Citation>Clin Microbiol Infect. 2020 Jun;26(6):773-779</Citation><ArticleIdList><ArticleId IdType="pubmed">32276116</ArticleId></ArticleIdList></Reference><Reference><Citation>N Engl J Med. 2020 May 28;382(22):2081-2090</Citation><ArticleIdList><ArticleId IdType="pubmed">32329971</ArticleId></ArticleIdList></Reference><Reference><Citation>J Clin Virol. 2020 Sep;130:104579</Citation><ArticleIdList><ArticleId IdType="pubmed">32795959</ArticleId></ArticleIdList></Reference><Reference><Citation>Clin Infect Dis. 2020 May 22;:</Citation><ArticleIdList><ArticleId IdType="pubmed">32442256</ArticleId></ArticleIdList></Reference><Reference><Citation>Radiology. 2020 Aug;296(2):E32-E40</Citation><ArticleIdList><ArticleId IdType="pubmed">32101510</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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