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Article: Improved Molecular Diagnosis of COVID-19 by the Novel, Highly Sensitive and Specific COVID-19-RdRp/Hel Real-Time Reverse Transcription-PCR Assay Validated In Vitro and with Clinical Specimens

TitleImproved Molecular Diagnosis of COVID-19 by the Novel, Highly Sensitive and Specific COVID-19-RdRp/Hel Real-Time Reverse Transcription-PCR Assay Validated In Vitro and with Clinical Specimens
Authors
Keywordscoronavirus
COVID-19
diagnostic
emerging
PCR
Issue Date2020
PublisherAmerican Society for Microbiology. The Journal's web site is located at http://jcm.asm.org/
Citation
Journal of Clinical Microbiology, 2020, v. 58 n. 5, p. article no. e00310-20 How to Cite?
AbstractOn 31 December 2019, the World Health Organization was informed of a cluster of cases of pneumonia of unknown etiology in Wuhan, China. Subsequent investigations identified a novel coronavirus, now named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), from the affected patients. Highly sensitive and specific laboratory diagnostics are important for controlling the rapidly evolving SARS-CoV-2-associated coronavirus disease 2019 (COVID-19) epidemic. In this study, we developed and compared the performance of three novel real-time reverse transcription-PCR (RT-PCR) assays targeting the RNA-dependent RNA polymerase (RdRp)/helicase (Hel), spike (S), and nucleocapsid (N) genes of SARS-CoV-2 with that of the reported RdRp-P2 assay, which is used in >30 European laboratories. Among the three novel assays, the COVID-19-RdRp/Hel assay had the lowest limit of detection in vitro (1.8 50% tissue culture infective doses [TCID50]/ml with genomic RNA and 11.2 RNA copies/reaction with in vitro RNA transcripts). Among 273 specimens from 15 patients with laboratory-confirmed COVID-19 in Hong Kong, 77 (28.2%) were positive by both the COVID-19-RdRp/Hel and RdRp-P2 assays. The COVID-19-RdRp/Hel assay was positive for an additional 42 RdRp-P2-negative specimens (119/273 [43.6%] versus 77/273 [28.2%]; P < 0.001), including 29/120 (24.2%) respiratory tract specimens and 13/153 (8.5%) non-respiratory tract specimens. The mean viral load of these specimens was 3.21 × 104 RNA copies/ml (range, 2.21 × 102 to 4.71 × 105 RNA copies/ml). The COVID-19-RdRp/Hel assay did not cross-react with other human-pathogenic coronaviruses and respiratory pathogens in cell culture and clinical specimens, whereas the RdRp-P2 assay cross-reacted with SARS-CoV in cell culture. The highly sensitive and specific COVID-19-RdRp/Hel assay may help to improve the laboratory diagnosis of COVID-19.
Persistent Identifierhttp://hdl.handle.net/10722/285241
ISSN
2019 Impact Factor: 5.897
2015 SCImago Journal Rankings: 2.151
PubMed Central ID

 

DC FieldValueLanguage
dc.contributor.authorChan, JFW-
dc.contributor.authorYip, CCY-
dc.contributor.authorTo, KKW-
dc.contributor.authorTang, THC-
dc.contributor.authorWong, SCY-
dc.contributor.authorLeung, KH-
dc.contributor.authorFung, AYF-
dc.contributor.authorNg, ACK-
dc.contributor.authorZou, Z-
dc.contributor.authorTsoi, HW-
dc.contributor.authorChoi, GKY-
dc.contributor.authorTam, AR-
dc.contributor.authorCheng, VCC-
dc.contributor.authorChan, KH-
dc.contributor.authorTsang, OTY-
dc.contributor.authorYuen, KY-
dc.date.accessioned2020-08-18T03:51:35Z-
dc.date.available2020-08-18T03:51:35Z-
dc.date.issued2020-
dc.identifier.citationJournal of Clinical Microbiology, 2020, v. 58 n. 5, p. article no. e00310-20-
dc.identifier.issn0095-1137-
dc.identifier.urihttp://hdl.handle.net/10722/285241-
dc.description.abstractOn 31 December 2019, the World Health Organization was informed of a cluster of cases of pneumonia of unknown etiology in Wuhan, China. Subsequent investigations identified a novel coronavirus, now named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), from the affected patients. Highly sensitive and specific laboratory diagnostics are important for controlling the rapidly evolving SARS-CoV-2-associated coronavirus disease 2019 (COVID-19) epidemic. In this study, we developed and compared the performance of three novel real-time reverse transcription-PCR (RT-PCR) assays targeting the RNA-dependent RNA polymerase (RdRp)/helicase (Hel), spike (S), and nucleocapsid (N) genes of SARS-CoV-2 with that of the reported RdRp-P2 assay, which is used in >30 European laboratories. Among the three novel assays, the COVID-19-RdRp/Hel assay had the lowest limit of detection in vitro (1.8 50% tissue culture infective doses [TCID50]/ml with genomic RNA and 11.2 RNA copies/reaction with in vitro RNA transcripts). Among 273 specimens from 15 patients with laboratory-confirmed COVID-19 in Hong Kong, 77 (28.2%) were positive by both the COVID-19-RdRp/Hel and RdRp-P2 assays. The COVID-19-RdRp/Hel assay was positive for an additional 42 RdRp-P2-negative specimens (119/273 [43.6%] versus 77/273 [28.2%]; P < 0.001), including 29/120 (24.2%) respiratory tract specimens and 13/153 (8.5%) non-respiratory tract specimens. The mean viral load of these specimens was 3.21 × 104 RNA copies/ml (range, 2.21 × 102 to 4.71 × 105 RNA copies/ml). The COVID-19-RdRp/Hel assay did not cross-react with other human-pathogenic coronaviruses and respiratory pathogens in cell culture and clinical specimens, whereas the RdRp-P2 assay cross-reacted with SARS-CoV in cell culture. The highly sensitive and specific COVID-19-RdRp/Hel assay may help to improve the laboratory diagnosis of COVID-19.-
dc.languageeng-
dc.publisherAmerican Society for Microbiology. The Journal's web site is located at http://jcm.asm.org/-
dc.relation.ispartofJournal of Clinical Microbiology-
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.-
dc.subjectcoronavirus-
dc.subjectCOVID-19-
dc.subjectdiagnostic-
dc.subjectemerging-
dc.subjectPCR-
dc.titleImproved Molecular Diagnosis of COVID-19 by the Novel, Highly Sensitive and Specific COVID-19-RdRp/Hel Real-Time Reverse Transcription-PCR Assay Validated In Vitro and with Clinical Specimens-
dc.typeArticle-
dc.identifier.emailChan, JFW: jfwchan@hku.hk-
dc.identifier.emailYip, CCY: yipcyril@hku.hk-
dc.identifier.emailTo, KKW: kelvinto@hku.hk-
dc.identifier.emailTang, THC: tanghct@hku.hk-
dc.identifier.emailWong, SCY: wcy288@hku.hk-
dc.identifier.emailLeung, KH: khl17@hku.hk-
dc.identifier.emailFung, AYF: agnes_fung@hku.hk-
dc.identifier.emailNg, ACK: nck912@hku.hk-
dc.identifier.emailTsoi, HW: hwtsoi@hkucc.hku.hk-
dc.identifier.emailCheng, VCC: vcccheng@hkucc.hku.hk-
dc.identifier.emailChan, KH: chankh2@hkucc.hku.hk-
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hk-
dc.identifier.authorityChan, JFW=rp01736-
dc.identifier.authorityYip, CCY=rp01721-
dc.identifier.authorityTo, KKW=rp01384-
dc.identifier.authorityTsoi, HW=rp00439-
dc.identifier.authorityChan, KH=rp01921-
dc.identifier.authorityYuen, KY=rp00366-
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1128/JCM.00310-20-
dc.identifier.pmid32132196-
dc.identifier.pmcidPMC7180250-
dc.identifier.scopuseid_2-s2.0-85083226778-
dc.identifier.hkuros312851-
dc.identifier.volume58-
dc.identifier.issue5-
dc.identifier.spagearticle no. e00310-
dc.identifier.epage20-
dc.publisher.placeUnited States-

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