Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2.
Identifieur interne : 000025 ( PubMed/Curation ); précédent : 000024; suivant : 000026Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2.
Auteurs : Yun Hee Baek [Corée du Sud] ; Jihye Um [Corée du Sud] ; Khristine Joy C. Antigua [Corée du Sud] ; Ji-Hyun Park [Corée du Sud] ; Yeonjae Kim [Corée du Sud] ; Sol Oh [Corée du Sud] ; Young-Il Kim [Corée du Sud] ; Won-Suk Choi [Corée du Sud] ; Seong Gyu Kim [Corée du Sud] ; Ju Hwan Jeong [Corée du Sud] ; Bum Sik Chin [Corée du Sud] ; Halcyon Dawn G. Nicolas [Corée du Sud] ; Ji-Young Ahn [Corée du Sud] ; Kyeong Seob Shin [Corée du Sud] ; Young Ki Choi [Corée du Sud] ; Jun-Sun Park [Corée du Sud] ; Min-Suk Song [Corée du Sud]Source :
- Emerging microbes & infections [ 2222-1751 ] ; 2020.
Abstract
The previous outbreaks of SARS-CoV and MERS-CoV have led researchers to study the role of diagnostics in impediment of further spread and transmission. With the recent emergence of the novel SARS-CoV-2, the availability of rapid, sensitive, and reliable diagnostic methods is essential for disease control. Hence, we have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the specific detection of SARS-CoV-2. The primer sets for RT-LAMP assay were designed to target the nucleocapsid gene of the viral RNA, and displayed a detection limit of 102 RNA copies close to that of qRT-PCR. Notably, the assay has exhibited a rapid detection span of 30 min combined with the colorimetric visualization. This test can detect specifically viral RNAs of the SARS-CoV-2 with no cross-reactivity to related coronaviruses, such as HCoV-229E, HCoV-NL63, HCoV-OC43, and MERS-CoV as well as human infectious influenza viruses (type B, H1N1pdm, H3N2, H5N1, H5N6, H5N8, and H7N9), and other respiratory disease-causing viruses (RSVA, RSVB, ADV, PIV, MPV, and HRV). Furthermore, the developed RT-LAMP assay has been evaluated using specimens collected from COVID-19 patients that exhibited high agreement to the qRT-PCR. Our RT-LAMP assay is simple to perform, less expensive, time-efficient, and can be used in clinical laboratories for preliminary detection of SARS-CoV-2 in suspected patients. In addition to the high sensitivity and specificity, this isothermal amplification conjugated with a single-tube colorimetric detection method may contribute to the public health responses and disease control, especially in the areas with limited laboratory capacities.
DOI: 10.1080/22221751.2020.1756698
PubMed: 32306853
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Antigua</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Park, Ji Hyun" sort="Park, Ji Hyun" uniqKey="Park J" first="Ji-Hyun" last="Park">Ji-Hyun Park</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Kim, Yeonjae" sort="Kim, Yeonjae" uniqKey="Kim Y" first="Yeonjae" last="Kim">Yeonjae Kim</name><affiliation wicri:level="1"><nlm:affiliation>Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul, Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Oh, Sol" sort="Oh, Sol" uniqKey="Oh S" first="Sol" last="Oh">Sol Oh</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Kim, Young Il" sort="Kim, Young Il" uniqKey="Kim Y" first="Young-Il" last="Kim">Young-Il Kim</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Choi, Won Suk" sort="Choi, Won Suk" uniqKey="Choi W" first="Won-Suk" last="Choi">Won-Suk Choi</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Kim, Seong Gyu" sort="Kim, Seong Gyu" uniqKey="Kim S" first="Seong Gyu" last="Kim">Seong Gyu Kim</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Jeong, Ju Hwan" sort="Jeong, Ju Hwan" uniqKey="Jeong J" first="Ju Hwan" last="Jeong">Ju Hwan Jeong</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Chin, Bum Sik" sort="Chin, Bum Sik" uniqKey="Chin B" first="Bum Sik" last="Chin">Bum Sik Chin</name><affiliation wicri:level="1"><nlm:affiliation>Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul, Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Nicolas, Halcyon Dawn G" sort="Nicolas, Halcyon Dawn G" uniqKey="Nicolas H" first="Halcyon Dawn G" last="Nicolas">Halcyon Dawn G. Nicolas</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Ahn, Ji Young" sort="Ahn, Ji Young" uniqKey="Ahn J" first="Ji-Young" last="Ahn">Ji-Young Ahn</name><affiliation wicri:level="1"><nlm:affiliation>School of Biological Sciences, Chungbuk National University, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>School of Biological Sciences, Chungbuk National University, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Shin, Kyeong Seob" sort="Shin, Kyeong Seob" uniqKey="Shin K" first="Kyeong Seob" last="Shin">Kyeong Seob Shin</name><affiliation wicri:level="1"><nlm:affiliation>Departments of Laboratory Medicine, Chungbuk National University College of Medicine, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Departments of Laboratory Medicine, Chungbuk National University College of Medicine, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Choi, Young Ki" sort="Choi, Young Ki" uniqKey="Choi Y" first="Young Ki" last="Choi">Young Ki Choi</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author><author><name sortKey="Park, Jun Sun" sort="Park, Jun Sun" uniqKey="Park J" first="Jun-Sun" last="Park">Jun-Sun Park</name><affiliation wicri:level="1"><nlm:affiliation>Research Institute of Public Health, National Medical Center, Seoul, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Research Institute of Public Health, National Medical Center, Seoul</wicri:regionArea></affiliation></author><author><name sortKey="Song, Min Suk" sort="Song, Min Suk" uniqKey="Song M" first="Min-Suk" last="Song">Min-Suk Song</name><affiliation wicri:level="1"><nlm:affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju</wicri:regionArea></affiliation></author></analytic><series><title level="j">Emerging microbes & infections</title><idno type="eISSN">2222-1751</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The previous outbreaks of SARS-CoV and MERS-CoV have led researchers to study the role of diagnostics in impediment of further spread and transmission. With the recent emergence of the novel SARS-CoV-2, the availability of rapid, sensitive, and reliable diagnostic methods is essential for disease control. Hence, we have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the specific detection of SARS-CoV-2. The primer sets for RT-LAMP assay were designed to target the nucleocapsid gene of the viral RNA, and displayed a detection limit of 10<sup>2</sup> RNA copies close to that of qRT-PCR. Notably, the assay has exhibited a rapid detection span of 30 min combined with the colorimetric visualization. This test can detect specifically viral RNAs of the SARS-CoV-2 with no cross-reactivity to related coronaviruses, such as HCoV-229E, HCoV-NL63, HCoV-OC43, and MERS-CoV as well as human infectious influenza viruses (type B, H1N1pdm, H3N2, H5N1, H5N6, H5N8, and H7N9), and other respiratory disease-causing viruses (RSVA, RSVB, ADV, PIV, MPV, and HRV). Furthermore, the developed RT-LAMP assay has been evaluated using specimens collected from COVID-19 patients that exhibited high agreement to the qRT-PCR. Our RT-LAMP assay is simple to perform, less expensive, time-efficient, and can be used in clinical laboratories for preliminary detection of SARS-CoV-2 in suspected patients. In addition to the high sensitivity and specificity, this isothermal amplification conjugated with a single-tube colorimetric detection method may contribute to the public health responses and disease control, especially in the areas with limited laboratory capacities.</div></front></TEI><pubmed><MedlineCitation Status="Publisher" Owner="NLM"><PMID Version="1">32306853</PMID><DateRevised><Year>2020</Year><Month>04</Month><Day>20</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">2222-1751</ISSN><JournalIssue CitedMedium="Internet"><PubDate><Year>2020</Year><Month>Apr</Month><Day>20</Day></PubDate></JournalIssue><Title>Emerging microbes & infections</Title><ISOAbbreviation>Emerg Microbes Infect</ISOAbbreviation></Journal><ArticleTitle>Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2.</ArticleTitle><Pagination><MedlinePgn>1-31</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1080/22221751.2020.1756698</ELocationID><Abstract><AbstractText>The previous outbreaks of SARS-CoV and MERS-CoV have led researchers to study the role of diagnostics in impediment of further spread and transmission. With the recent emergence of the novel SARS-CoV-2, the availability of rapid, sensitive, and reliable diagnostic methods is essential for disease control. Hence, we have developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the specific detection of SARS-CoV-2. The primer sets for RT-LAMP assay were designed to target the nucleocapsid gene of the viral RNA, and displayed a detection limit of 10<sup>2</sup> RNA copies close to that of qRT-PCR. Notably, the assay has exhibited a rapid detection span of 30 min combined with the colorimetric visualization. This test can detect specifically viral RNAs of the SARS-CoV-2 with no cross-reactivity to related coronaviruses, such as HCoV-229E, HCoV-NL63, HCoV-OC43, and MERS-CoV as well as human infectious influenza viruses (type B, H1N1pdm, H3N2, H5N1, H5N6, H5N8, and H7N9), and other respiratory disease-causing viruses (RSVA, RSVB, ADV, PIV, MPV, and HRV). Furthermore, the developed RT-LAMP assay has been evaluated using specimens collected from COVID-19 patients that exhibited high agreement to the qRT-PCR. Our RT-LAMP assay is simple to perform, less expensive, time-efficient, and can be used in clinical laboratories for preliminary detection of SARS-CoV-2 in suspected patients. In addition to the high sensitivity and specificity, this isothermal amplification conjugated with a single-tube colorimetric detection method may contribute to the public health responses and disease control, especially in the areas with limited laboratory capacities.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Baek</LastName><ForeName>Yun Hee</ForeName><Initials>YH</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Um</LastName><ForeName>Jihye</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Research Institute of Public Health, National Medical Center, Seoul, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Antigua</LastName><ForeName>Khristine Joy C</ForeName><Initials>KJC</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Park</LastName><ForeName>Ji-Hyun</ForeName><Initials>JH</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kim</LastName><ForeName>Yeonjae</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul, Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Oh</LastName><ForeName>Sol</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kim</LastName><ForeName>Young-Il</ForeName><Initials>YI</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Choi</LastName><ForeName>Won-Suk</ForeName><Initials>WS</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kim</LastName><ForeName>Seong Gyu</ForeName><Initials>SG</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jeong</LastName><ForeName>Ju Hwan</ForeName><Initials>JH</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Chin</LastName><ForeName>Bum Sik</ForeName><Initials>BS</Initials><AffiliationInfo><Affiliation>Center for Infectious Diseases Research, Department of Internal Medicine, National Medical Center, Seoul, Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Nicolas</LastName><ForeName>Halcyon Dawn G</ForeName><Initials>HDG</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ahn</LastName><ForeName>Ji-Young</ForeName><Initials>JY</Initials><AffiliationInfo><Affiliation>School of Biological Sciences, Chungbuk National University, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Shin</LastName><ForeName>Kyeong Seob</ForeName><Initials>KS</Initials><AffiliationInfo><Affiliation>Departments of Laboratory Medicine, Chungbuk National University College of Medicine, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Choi</LastName><ForeName>Young Ki</ForeName><Initials>YK</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Park</LastName><ForeName>Jun-Sun</ForeName><Initials>JS</Initials><AffiliationInfo><Affiliation>Research Institute of Public Health, National Medical Center, Seoul, Republic of Korea.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Song</LastName><ForeName>Min-Suk</ForeName><Initials>MS</Initials><AffiliationInfo><Affiliation>Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute, Cheongju, Republic of Korea.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>04</Month><Day>20</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Emerg Microbes Infect</MedlineTA><NlmUniqueID>101594885</NlmUniqueID><ISSNLinking>2222-1751</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">COVID-19</Keyword><Keyword MajorTopicYN="N">SARS-CoV-2</Keyword><Keyword MajorTopicYN="N">colorimetric detection</Keyword><Keyword MajorTopicYN="N">molecular diagnosis</Keyword><Keyword MajorTopicYN="N">novel coronavirus</Keyword><Keyword MajorTopicYN="N">reverse transcription-loop-mediated isothermal amplification</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>4</Month><Day>21</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>4</Month><Day>21</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>4</Month><Day>21</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>aheadofprint</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">32306853</ArticleId><ArticleId IdType="doi">10.1080/22221751.2020.1756698</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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