A Simple and Multiplex Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of SARS-CoV.
Identifieur interne : 000936 ( PubMed/Checkpoint ); précédent : 000935; suivant : 000937A Simple and Multiplex Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of SARS-CoV.
Auteurs : Jin Hwa Kim [Corée du Sud] ; Minhee Kang [Corée du Sud] ; Eunkyoung Park [Corée du Sud] ; Doo Ryeon Chung [Corée du Sud] ; Jiyeon Kim [Corée du Sud] ; Eung Soo Hwang [Corée du Sud]Source :
- Biochip journal [ 1976-0280 ] ; 2019.
Abstract
The current diagnosis of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) relies on laboratory-based tests since its clinical features are nonspecific, unlike other respiratory pathogens. Therefore, the development of a rapid and simple method for on-site detection of SARS-CoV is crucial for the identification and prevention of future SARS outbreaks. In this study, a simple colorimetric and multiplex loop-mediated isothermal amplification (LAMP) assay was developed to rapid screening of severe acute respiratory syndrome-associated coronavirus (SARS-CoV). It can be visually detected based on color change and monitored in real-time with fluorescent signals. The performance of this assay, based on six primers targeting open reading frame (ORF1b) and nucleocapsid (N) genes located in different regions of the SARS-CoV, was compared with real-time RT-PCR assay using various concentrations of target genes. The detection limit of the LAMP assay was comparable to that of real-time RT-PCR assay and therefore a few target RNA to 104-105 copies could be detected within a short period of time (20-25 min). In addition, we established a multiplex real-time LAMP assay to simultaneously detect two target regions within the SARS-CoV genome. Two target sequences were amplified by specific primers in the same reaction tube and revealed that it was able to detect down to 105 copies. The standard curve had a linear relationship with similar amplification efficiencies. The LAMP assay results in shorter "sample-to-answer" time than conventional PCR method. Therefore, it is suitable not only for diagnosis of clinical test, but also for surveillance of SARS virus in developing countries.
DOI: 10.1007/s13206-019-3404-3
PubMed: 32226589
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Minhee" uniqKey="Kang M" first="Minhee" last="Kang">Minhee Kang</name><affiliation wicri:level="3"><nlm:affiliation>1Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>1Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Park, Eunkyoung" sort="Park, Eunkyoung" uniqKey="Park E" first="Eunkyoung" last="Park">Eunkyoung Park</name><affiliation wicri:level="3"><nlm:affiliation>1Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>1Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Chung, Doo Ryeon" sort="Chung, Doo Ryeon" uniqKey="Chung D" first="Doo Ryeon" last="Chung">Doo Ryeon Chung</name><affiliation wicri:level="3"><nlm:affiliation>3Center for Infection Prevention and Control, Samsung Medical Center, Seoul, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>3Center for Infection Prevention and Control, Samsung Medical Center, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Kim, Jiyeon" sort="Kim, Jiyeon" uniqKey="Kim J" first="Jiyeon" last="Kim">Jiyeon Kim</name><affiliation wicri:level="3"><nlm:affiliation>6Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, South Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>6Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author><author><name sortKey="Hwang, Eung Soo" sort="Hwang, Eung Soo" uniqKey="Hwang E" first="Eung Soo" last="Hwang">Eung Soo Hwang</name><affiliation wicri:level="3"><nlm:affiliation>6Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, South Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>6Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de Séoul</region></placeName></affiliation></author></analytic><series><title level="j">Biochip journal</title><idno type="ISSN">1976-0280</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The current diagnosis of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) relies on laboratory-based tests since its clinical features are nonspecific, unlike other respiratory pathogens. Therefore, the development of a rapid and simple method for on-site detection of SARS-CoV is crucial for the identification and prevention of future SARS outbreaks. In this study, a simple colorimetric and multiplex loop-mediated isothermal amplification (LAMP) assay was developed to rapid screening of severe acute respiratory syndrome-associated coronavirus (SARS-CoV). It can be visually detected based on color change and monitored in real-time with fluorescent signals. The performance of this assay, based on six primers targeting open reading frame (ORF1b) and nucleocapsid (N) genes located in different regions of the SARS-CoV, was compared with real-time RT-PCR assay using various concentrations of target genes. The detection limit of the LAMP assay was comparable to that of real-time RT-PCR assay and therefore a few target RNA to 10<sup>4</sup>-10<sup>5</sup> copies could be detected within a short period of time (20-25 min). In addition, we established a multiplex real-time LAMP assay to simultaneously detect two target regions within the SARS-CoV genome. Two target sequences were amplified by specific primers in the same reaction tube and revealed that it was able to detect down to 10<sup>5</sup> copies. The standard curve had a linear relationship with similar amplification efficiencies. The LAMP assay results in shorter "sample-to-answer" time than conventional PCR method. Therefore, it is suitable not only for diagnosis of clinical test, but also for surveillance of SARS virus in developing countries.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">32226589</PMID><DateRevised><Year>2020</Year><Month>04</Month><Day>03</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">1976-0280</ISSN><JournalIssue CitedMedium="Print"><Volume>13</Volume><Issue>4</Issue><PubDate><Year>2019</Year></PubDate></JournalIssue><Title>Biochip journal</Title><ISOAbbreviation>Biochip J</ISOAbbreviation></Journal><ArticleTitle>A Simple and Multiplex Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of SARS-CoV.</ArticleTitle><Pagination><MedlinePgn>341-351</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s13206-019-3404-3</ELocationID><Abstract><AbstractText>The current diagnosis of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) relies on laboratory-based tests since its clinical features are nonspecific, unlike other respiratory pathogens. Therefore, the development of a rapid and simple method for on-site detection of SARS-CoV is crucial for the identification and prevention of future SARS outbreaks. In this study, a simple colorimetric and multiplex loop-mediated isothermal amplification (LAMP) assay was developed to rapid screening of severe acute respiratory syndrome-associated coronavirus (SARS-CoV). It can be visually detected based on color change and monitored in real-time with fluorescent signals. The performance of this assay, based on six primers targeting open reading frame (ORF1b) and nucleocapsid (N) genes located in different regions of the SARS-CoV, was compared with real-time RT-PCR assay using various concentrations of target genes. The detection limit of the LAMP assay was comparable to that of real-time RT-PCR assay and therefore a few target RNA to 10<sup>4</sup>-10<sup>5</sup> copies could be detected within a short period of time (20-25 min). In addition, we established a multiplex real-time LAMP assay to simultaneously detect two target regions within the SARS-CoV genome. Two target sequences were amplified by specific primers in the same reaction tube and revealed that it was able to detect down to 10<sup>5</sup> copies. The standard curve had a linear relationship with similar amplification efficiencies. The LAMP assay results in shorter "sample-to-answer" time than conventional PCR method. Therefore, it is suitable not only for diagnosis of clinical test, but also for surveillance of SARS virus in developing countries.</AbstractText><AbstractText Label="Electronic Supplementary Material" NlmCategory="UNASSIGNED">Supplementary material is available for this article at 10.1007/s13206-019-3404-3 and is accessible for authorized users.</AbstractText><CopyrightInformation>© The Korean BioChip Society and Springer 2019.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y" EqualContrib="Y"><LastName>Kim</LastName><ForeName>Jin Hwa</ForeName><Initials>JH</Initials><AffiliationInfo><Affiliation>1Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.</Affiliation><Identifier Source="GRID">grid.264381.a</Identifier><Identifier Source="ISNI">0000 0001 2181 989X</Identifier></AffiliationInfo></Author><Author ValidYN="Y" EqualContrib="Y"><LastName>Kang</LastName><ForeName>Minhee</ForeName><Initials>M</Initials><Identifier Source="ORCID">0000-0003-0330-7828</Identifier><AffiliationInfo><Affiliation>1Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.</Affiliation><Identifier Source="GRID">grid.264381.a</Identifier><Identifier Source="ISNI">0000 0001 2181 989X</Identifier></AffiliationInfo><AffiliationInfo><Affiliation>2Department of Medical Device Management and Research, SAIHST, Sungkyunkwan University, Seoul, Republic of Korea.</Affiliation><Identifier Source="GRID">grid.264381.a</Identifier><Identifier Source="ISNI">0000 0001 2181 989X</Identifier></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Park</LastName><ForeName>Eunkyoung</ForeName><Initials>E</Initials><AffiliationInfo><Affiliation>1Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.</Affiliation><Identifier Source="GRID">grid.264381.a</Identifier><Identifier Source="ISNI">0000 0001 2181 989X</Identifier></AffiliationInfo><AffiliationInfo><Affiliation>2Department of Medical Device Management and Research, SAIHST, Sungkyunkwan University, Seoul, Republic of Korea.</Affiliation><Identifier Source="GRID">grid.264381.a</Identifier><Identifier Source="ISNI">0000 0001 2181 989X</Identifier></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Chung</LastName><ForeName>Doo Ryeon</ForeName><Initials>DR</Initials><Identifier Source="ORCID">0000-0001-9267-101X</Identifier><AffiliationInfo><Affiliation>3Center for Infection Prevention and Control, Samsung Medical Center, Seoul, Republic of Korea.</Affiliation><Identifier Source="GRID">grid.414964.a</Identifier><Identifier Source="ISNI">0000 0001 0640 5613</Identifier></AffiliationInfo><AffiliationInfo><Affiliation>Asia Pacific Foundation for Infectious Diseases (APFID), Seoul, Republic of Korea.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>5Division of Infectious Diseases, Department of Internal Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.</Affiliation><Identifier Source="GRID">grid.264381.a</Identifier><Identifier Source="ISNI">0000 0001 2181 989X</Identifier></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kim</LastName><ForeName>Jiyeon</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>6Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, South Korea.</Affiliation><Identifier Source="GRID">grid.31501.36</Identifier><Identifier Source="ISNI">0000 0004 0470 5905</Identifier></AffiliationInfo><AffiliationInfo><Affiliation>7Institute of Endemic Diseases, Seoul National University Medical Research Center, Seoul, South Korea.</Affiliation><Identifier Source="GRID">grid.412484.f</Identifier><Identifier Source="ISNI">0000 0001 0302 820X</Identifier></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hwang</LastName><ForeName>Eung Soo</ForeName><Initials>ES</Initials><AffiliationInfo><Affiliation>6Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, South Korea.</Affiliation><Identifier Source="GRID">grid.31501.36</Identifier><Identifier Source="ISNI">0000 0004 0470 5905</Identifier></AffiliationInfo><AffiliationInfo><Affiliation>7Institute of Endemic Diseases, Seoul National University Medical Research Center, Seoul, South Korea.</Affiliation><Identifier Source="GRID">grid.412484.f</Identifier><Identifier Source="ISNI">0000 0001 0302 820X</Identifier></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>11</Month><Day>11</Day></ArticleDate></Article><MedlineJournalInfo><Country>Korea (South)</Country><MedlineTA>Biochip J</MedlineTA><NlmUniqueID>101517318</NlmUniqueID><ISSNLinking>1976-0280</ISSNLinking></MedlineJournalInfo><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Colorimetric detection</Keyword><Keyword MajorTopicYN="N">Loop-mediated isothermal amplification</Keyword><Keyword MajorTopicYN="N">Point-of-care test</Keyword><Keyword MajorTopicYN="N">SARS-CoV</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>07</Month><Day>15</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>08</Month><Day>23</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>4</Month><Day>1</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>1</Month><Day>1</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2019</Year><Month>1</Month><Day>1</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">32226589</ArticleId><ArticleId IdType="doi">10.1007/s13206-019-3404-3</ArticleId><ArticleId IdType="pii">3404</ArticleId><ArticleId IdType="pmc">PMC7097549</ArticleId></ArticleIdList><pmc-dir>pmcsd</pmc-dir>
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Kang</name><name sortKey="Kim, Jiyeon" sort="Kim, Jiyeon" uniqKey="Kim J" first="Jiyeon" last="Kim">Jiyeon Kim</name><name sortKey="Park, Eunkyoung" sort="Park, Eunkyoung" uniqKey="Park E" first="Eunkyoung" last="Park">Eunkyoung Park</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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