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Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus.

Identifieur interne : 000B99 ( PubMed/Curation ); précédent : 000B98; suivant : 000C00

Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus.

Auteurs : Yun Young Go [Corée du Sud] ; Yeon-Sook Kim [Corée du Sud] ; Shinhye Cheon [Corée du Sud] ; Sangwoo Nam [Corée du Sud] ; Keun Bon Ku [Corée du Sud] ; Meehyein Kim [Corée du Sud] ; Nam Hyuk Cho [Corée du Sud] ; Hyun Park [Corée du Sud] ; Pei-Yu Alison Lee [États-Unis] ; Yu-Chun Lin [États-Unis] ; Yun-Long Tsai [États-Unis] ; Hwa-Tang Thomas Wang [États-Unis] ; Udeni B R. Balasuriya [États-Unis]

Source :

The Journal of molecular diagnostics : JMD [ 1943-7811 ] ; 2017.

RBID : pubmed:28807812

Descripteurs français

KwdFr :
- ARN viral (génétique), ARN viral (isolement et purification), Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Coronavirus du syndrome respiratoire du Moyen-Orient (pathogénicité), Humains, Infections à coronavirus (diagnostic), Infections à coronavirus (génétique), Infections à coronavirus (virologie), Réaction de polymérisation en chaine en temps réel (), République de Corée (épidémiologie), Transcription inverse (génétique).
MESH :
- diagnostic : Infections à coronavirus.
- génétique : ARN viral, Coronavirus du syndrome respiratoire du Moyen-Orient, Infections à coronavirus, Transcription inverse.
- isolement et purification : ARN viral, Coronavirus du syndrome respiratoire du Moyen-Orient.
- pathogénicité : Coronavirus du syndrome respiratoire du Moyen-Orient.
- virologie : Infections à coronavirus.
- épidémiologie : République de Corée.
- Humains, Réaction de polymérisation en chaine en temps réel.

English descriptors

KwdEn :
- Coronavirus Infections (diagnosis), Coronavirus Infections (genetics), Coronavirus Infections (virology), Humans, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), Middle East Respiratory Syndrome Coronavirus (pathogenicity), RNA, Viral (genetics), RNA, Viral (isolation & purification), Real-Time Polymerase Chain Reaction (methods), Republic of Korea (epidemiology), Reverse Transcription (genetics).
MESH :
- chemical , genetics : RNA, Viral.
- diagnosis : Coronavirus Infections.
- epidemiology : Republic of Korea.
- genetics : Coronavirus Infections, Middle East Respiratory Syndrome Coronavirus, Reverse Transcription.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus, RNA, Viral.
- methods : Real-Time Polymerase Chain Reaction.
- pathogenicity : Middle East Respiratory Syndrome Coronavirus.
- virology : Coronavirus Infections.
- Humans.

Abstract

Middle East respiratory syndrome (MERS) is an emerging zoonotic viral respiratory disease that was first identified in Saudi Arabia in 2012. In 2015, the largest MERS outbreak outside of the Middle East region occurred in the Republic of Korea. The rapid nosocomial transmission of MERS-coronavirus (MERS-CoV) in Korean health care settings highlighted the importance and urgent need for a rapid and reliable on-site diagnostic assay to implement effective control and preventive measures. Here, the evaluation and validation of two newly developed reverse transcription-insulated isothermal PCR (RT-iiPCR) methods targeting the ORF1a and upE genes of MERS-CoV are described. Compared with World Health Organization-recommended singleplex real-time quantitative RT-PCR (RT-qPCR) assays, both RT-iiPCR assays had comparable analytical sensitivity for the detection of MERS-CoV RNA in tissue culture fluid and in sputum samples spiked with infectious virus. Furthermore, clinical evaluation was performed with sputum samples collected from subjects with acute and chronic respiratory illnesses, including patients infected with MERS-CoV. The overall agreement values between the two RT-iiPCR assays and the reference RT-qPCR assays were 98.06% (95% CI, 94.43%-100%; κ = 0.96) and 99.03% (95% CI, 95.88%-100%; κ = 0.99) for ORF1a and upE assays, respectively. The ORF1a and upE MERS-CoV RT-iiPCR assays coupled with a field-deployable system provide a platform for a highly sensitive and specific on-site tool for diagnosis of MERS-CoV infections.

DOI: 10.1016/j.jmoldx.2017.06.007
PubMed: 28807812

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<term>Humans</term>
<term>Middle East Respiratory Syndrome Coronavirus (genetics)</term>
<term>Middle East Respiratory Syndrome Coronavirus (isolation & purification)</term>
<term>Middle East Respiratory Syndrome Coronavirus (pathogenicity)</term>
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<term>Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification)</term>
<term>Coronavirus du syndrome respiratoire du Moyen-Orient (pathogénicité)</term>
<term>Humains</term>
<term>Infections à coronavirus (diagnostic)</term>
<term>Infections à coronavirus (génétique)</term>
<term>Infections à coronavirus (virologie)</term>
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<term>Middle East Respiratory Syndrome Coronavirus</term>
<term>Reverse Transcription</term>
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<term>Coronavirus du syndrome respiratoire du Moyen-Orient</term>
<term>Infections à coronavirus</term>
<term>Transcription inverse</term>
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<front><div type="abstract" xml:lang="en">Middle East respiratory syndrome (MERS) is an emerging zoonotic viral respiratory disease that was first identified in Saudi Arabia in 2012. In 2015, the largest MERS outbreak outside of the Middle East region occurred in the Republic of Korea. The rapid nosocomial transmission of MERS-coronavirus (MERS-CoV) in Korean health care settings highlighted the importance and urgent need for a rapid and reliable on-site diagnostic assay to implement effective control and preventive measures. Here, the evaluation and validation of two newly developed reverse transcription-insulated isothermal PCR (RT-iiPCR) methods targeting the ORF1a and upE genes of MERS-CoV are described. Compared with World Health Organization-recommended singleplex real-time quantitative RT-PCR (RT-qPCR) assays, both RT-iiPCR assays had comparable analytical sensitivity for the detection of MERS-CoV RNA in tissue culture fluid and in sputum samples spiked with infectious virus. Furthermore, clinical evaluation was performed with sputum samples collected from subjects with acute and chronic respiratory illnesses, including patients infected with MERS-CoV. The overall agreement values between the two RT-iiPCR assays and the reference RT-qPCR assays were 98.06% (95% CI, 94.43%-100%; κ = 0.96) and 99.03% (95% CI, 95.88%-100%; κ = 0.99) for ORF1a and upE assays, respectively. The ORF1a and upE MERS-CoV RT-iiPCR assays coupled with a field-deployable system provide a platform for a highly sensitive and specific on-site tool for diagnosis of MERS-CoV infections.</div>
</front>
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<DateCompleted><Year>2018</Year>
<Month>06</Month>
<Day>06</Day>
</DateCompleted>
<DateRevised><Year>2020</Year>
<Month>04</Month>
<Day>02</Day>
</DateRevised>
<Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1943-7811</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>19</Volume>
<Issue>6</Issue>
<PubDate><Year>2017</Year>
<Month>11</Month>
</PubDate>
</JournalIssue>
<Title>The Journal of molecular diagnostics : JMD</Title>
<ISOAbbreviation>J Mol Diagn</ISOAbbreviation>
</Journal>
<ArticleTitle>Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus.</ArticleTitle>
<Pagination><MedlinePgn>817-827</MedlinePgn>
</Pagination>
<ELocationID EIdType="pii" ValidYN="Y">S1525-1578(17)30188-5</ELocationID>
<ELocationID EIdType="doi" ValidYN="Y">10.1016/j.jmoldx.2017.06.007</ELocationID>
<Abstract><AbstractText>Middle East respiratory syndrome (MERS) is an emerging zoonotic viral respiratory disease that was first identified in Saudi Arabia in 2012. In 2015, the largest MERS outbreak outside of the Middle East region occurred in the Republic of Korea. The rapid nosocomial transmission of MERS-coronavirus (MERS-CoV) in Korean health care settings highlighted the importance and urgent need for a rapid and reliable on-site diagnostic assay to implement effective control and preventive measures. Here, the evaluation and validation of two newly developed reverse transcription-insulated isothermal PCR (RT-iiPCR) methods targeting the ORF1a and upE genes of MERS-CoV are described. Compared with World Health Organization-recommended singleplex real-time quantitative RT-PCR (RT-qPCR) assays, both RT-iiPCR assays had comparable analytical sensitivity for the detection of MERS-CoV RNA in tissue culture fluid and in sputum samples spiked with infectious virus. Furthermore, clinical evaluation was performed with sputum samples collected from subjects with acute and chronic respiratory illnesses, including patients infected with MERS-CoV. The overall agreement values between the two RT-iiPCR assays and the reference RT-qPCR assays were 98.06% (95% CI, 94.43%-100%; κ = 0.96) and 99.03% (95% CI, 95.88%-100%; κ = 0.99) for ORF1a and upE assays, respectively. The ORF1a and upE MERS-CoV RT-iiPCR assays coupled with a field-deployable system provide a platform for a highly sensitive and specific on-site tool for diagnosis of MERS-CoV infections.</AbstractText>
<CopyrightInformation>Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.</CopyrightInformation>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Go</LastName>
<ForeName>Yun Young</ForeName>
<Initials>YY</Initials>
<AffiliationInfo><Affiliation>Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon, Republic of Korea. Electronic address: yygo@krict.re.kr.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Kim</LastName>
<ForeName>Yeon-Sook</ForeName>
<Initials>YS</Initials>
<AffiliationInfo><Affiliation>Department of Medicine, Chungnam National University School of Medicine, Daejeon, Republic of Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Cheon</LastName>
<ForeName>Shinhye</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>Department of Medicine, Chungnam National University School of Medicine, Daejeon, Republic of Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Nam</LastName>
<ForeName>Sangwoo</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon, Republic of Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Ku</LastName>
<ForeName>Keun Bon</ForeName>
<Initials>KB</Initials>
<AffiliationInfo><Affiliation>Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Kim</LastName>
<ForeName>Meehyein</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>Center for Virus Research and Testing, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon, Republic of Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Cho</LastName>
<ForeName>Nam Hyuk</ForeName>
<Initials>NH</Initials>
<AffiliationInfo><Affiliation>Department of Microbiology and Immunology, and Department of Biomedical Science, Seoul National University College of Medicine and Bundang Hospital, Seoul, Republic of Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Park</LastName>
<ForeName>Hyun</ForeName>
<Initials>H</Initials>
<AffiliationInfo><Affiliation>Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Republic of Korea.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Alison Lee</LastName>
<ForeName>Pei-Yu</ForeName>
<Initials>PY</Initials>
<AffiliationInfo><Affiliation>GeneReach USA, Lexington, Massachusetts.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Lin</LastName>
<ForeName>Yu-Chun</ForeName>
<Initials>YC</Initials>
<AffiliationInfo><Affiliation>GeneReach USA, Lexington, Massachusetts.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Tsai</LastName>
<ForeName>Yun-Long</ForeName>
<Initials>YL</Initials>
<AffiliationInfo><Affiliation>GeneReach USA, Lexington, Massachusetts.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Thomas Wang</LastName>
<ForeName>Hwa-Tang</ForeName>
<Initials>HT</Initials>
<AffiliationInfo><Affiliation>GeneReach USA, Lexington, Massachusetts.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Balasuriya</LastName>
<ForeName>Udeni B R</ForeName>
<Initials>UBR</Initials>
<AffiliationInfo><Affiliation>Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, College of Agriculture, Food and Environment and Department of Microbiology, Immunology and Molecular Genetics, College of Medicine, University of Kentucky, Lexington, Kentucky. Electronic address: ubalasuriya@uky.edu.</Affiliation>
</AffiliationInfo>
</Author>
</AuthorList>
<Language>eng</Language>
<PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType>
<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
</PublicationTypeList>
<ArticleDate DateType="Electronic"><Year>2017</Year>
<Month>08</Month>
<Day>12</Day>
</ArticleDate>
</Article>
<MedlineJournalInfo><Country>United States</Country>
<MedlineTA>J Mol Diagn</MedlineTA>
<NlmUniqueID>100893612</NlmUniqueID>
<ISSNLinking>1525-1578</ISSNLinking>
</MedlineJournalInfo>
<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D012367">RNA, Viral</NameOfSubstance>
</Chemical>
</ChemicalList>
<CitationSubset>IM</CitationSubset>
<MeshHeadingList><MeshHeading><DescriptorName UI="D018352" MajorTopicYN="N">Coronavirus Infections</DescriptorName>
<QualifierName UI="Q000175" MajorTopicYN="Y">diagnosis</QualifierName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D065207" MajorTopicYN="N">Middle East Respiratory Syndrome Coronavirus</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName>
<QualifierName UI="Q000472" MajorTopicYN="N">pathogenicity</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012367" MajorTopicYN="N">RNA, Viral</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D060888" MajorTopicYN="N">Real-Time Polymerase Chain Reaction</DescriptorName>
<QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D056910" MajorTopicYN="N">Republic of Korea</DescriptorName>
<QualifierName UI="Q000453" MajorTopicYN="N">epidemiology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D048348" MajorTopicYN="N">Reverse Transcription</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
</MeshHeading>
</MeshHeadingList>
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Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus.

Evaluation and Clinical Validation of Two Field-Deployable Reverse Transcription-Insulated Isothermal PCR Assays for the Detection of the Middle East Respiratory Syndrome-Coronavirus.

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