Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19).
Identifieur interne : 000735 ( Main/Exploration ); précédent : 000734; suivant : 000736Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19).
Auteurs : Joungha Won [Corée du Sud] ; Solji Lee [Corée du Sud] ; Myungsun Park [Corée du Sud] ; Tai Young Kim [Corée du Sud] ; Mingu Gordon Park [Corée du Sud] ; Byung Yoon Choi [Corée du Sud] ; Dongwan Kim [Corée du Sud] ; Hyeshik Chang [Corée du Sud] ; V Narry Kim [Corée du Sud] ; C Justin Lee [Corée du Sud]Source :
- Experimental neurobiology [ 1226-2560 ] ; 2020.
Abstract
The severe acute respiratory coronavirus 2 (SARS-CoV-2), which emerged in December 2019 in Wuhan, China, has spread rapidly to over a dozen countries. Especially, the spike of case numbers in South Korea sparks pandemic worries. This virus is reported to spread mainly through personto- person contact via respiratory droplets generated by coughing and sneezing, or possibly through surface contaminated by people coughing or sneezing on them. More critically, there have been reports about the possibility of this virus to transmit even before a virus-carrying person to show symptoms. Therefore, a low-cost, easy-access protocol for early detection of this virus is desperately needed. Here, we have established a real-time reverse-transcription PCR (rtPCR)-based assay protocol composed of easy specimen self-collection from a subject via pharyngeal swab, Trizolbased RNA purification, and SYBR Green-based rtPCR. This protocol shows an accuracy and sensitivity limit of 1-10 virus particles as we tested with a known lentivirus. The cost for each sample is estimated to be less than 15 US dollars. Overall time it takes for an entire protocol is estimated to be less than 4 hours. We propose a cost-effective, quick-and-easy method for early detection of SARS-CoV-2 at any conventional Biosafety Level II laboratories that are equipped with a rtPCR machine. Our newly developed protocol should be helpful for a first-hand screening of the asymptomatic virus-carriers for further prevention of transmission and early intervention and treatment for the rapidly propagating virus.
DOI: 10.5607/en20009
PubMed: 32156101
Affiliations:
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uniqKey="Choi B" first="Byung Yoon" last="Choi">Byung Yoon Choi</name><affiliation wicri:level="1"><nlm:affiliation>Department of Otorhinolaryngology, Seoul National University Bundang Hospital, Seongnam 13620, Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Otorhinolaryngology, Seoul National University Bundang Hospital, Seongnam 13620</wicri:regionArea><wicri:noRegion>Seongnam 13620</wicri:noRegion></affiliation></author><author><name sortKey="Kim, Dongwan" sort="Kim, Dongwan" uniqKey="Kim D" first="Dongwan" last="Kim">Dongwan Kim</name><affiliation wicri:level="1"><nlm:affiliation>Center for RNA Research, Institute for Basic Science, Seoul 08826, Korea.</nlm:affiliation><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Center for RNA Research, Institute for Basic Science, Seoul 08826</wicri:regionArea><placeName><settlement type="city">Séoul</settlement><region type="capital">Région capitale de 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type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The severe acute respiratory coronavirus 2 (SARS-CoV-2), which emerged in December 2019 in Wuhan, China, has spread rapidly to over a dozen countries. Especially, the spike of case numbers in South Korea sparks pandemic worries. This virus is reported to spread mainly through personto- person contact via respiratory droplets generated by coughing and sneezing, or possibly through surface contaminated by people coughing or sneezing on them. More critically, there have been reports about the possibility of this virus to transmit even before a virus-carrying person to show symptoms. Therefore, a low-cost, easy-access protocol for early detection of this virus is desperately needed. Here, we have established a real-time reverse-transcription PCR (rtPCR)-based assay protocol composed of easy specimen self-collection from a subject via pharyngeal swab, Trizolbased RNA purification, and SYBR Green-based rtPCR. This protocol shows an accuracy and sensitivity limit of 1-10 virus particles as we tested with a known lentivirus. The cost for each sample is estimated to be less than 15 US dollars. Overall time it takes for an entire protocol is estimated to be less than 4 hours. We propose a cost-effective, quick-and-easy method for early detection of SARS-CoV-2 at any conventional Biosafety Level II laboratories that are equipped with a rtPCR machine. Our newly developed protocol should be helpful for a first-hand screening of the asymptomatic virus-carriers for further prevention of transmission and early intervention and treatment for the rapidly propagating virus.</div></front></TEI><affiliations><list><country><li>Corée du Sud</li></country><region><li>Région capitale de Séoul</li></region><settlement><li>Séoul</li></settlement></list><tree><country name="Corée du Sud"><noRegion><name sortKey="Won, Joungha" sort="Won, Joungha" uniqKey="Won J" first="Joungha" last="Won">Joungha Won</name></noRegion><name sortKey="Chang, Hyeshik" sort="Chang, Hyeshik" uniqKey="Chang H" first="Hyeshik" last="Chang">Hyeshik Chang</name><name sortKey="Choi, Byung Yoon" sort="Choi, Byung Yoon" uniqKey="Choi B" first="Byung Yoon" last="Choi">Byung Yoon Choi</name><name sortKey="Kim, Dongwan" sort="Kim, Dongwan" uniqKey="Kim D" first="Dongwan" last="Kim">Dongwan Kim</name><name sortKey="Kim, Tai Young" sort="Kim, Tai Young" uniqKey="Kim T" first="Tai Young" last="Kim">Tai Young Kim</name><name sortKey="Kim, V Narry" sort="Kim, V Narry" uniqKey="Kim V" first="V Narry" last="Kim">V Narry Kim</name><name sortKey="Lee, C Justin" sort="Lee, C Justin" uniqKey="Lee C" first="C Justin" last="Lee">C Justin Lee</name><name sortKey="Lee, Solji" sort="Lee, Solji" uniqKey="Lee S" first="Solji" last="Lee">Solji Lee</name><name sortKey="Park, Mingu Gordon" sort="Park, Mingu Gordon" uniqKey="Park M" first="Mingu Gordon" last="Park">Mingu Gordon Park</name><name sortKey="Park, Myungsun" sort="Park, Myungsun" uniqKey="Park M" first="Myungsun" last="Park">Myungsun Park</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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