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Article: Development and Evaluation of Novel and Highly Sensitive Single-Tube Nested Real-Time RT-PCR Assays for SARS-CoV-2 Detection

TitleDevelopment and Evaluation of Novel and Highly Sensitive Single-Tube Nested Real-Time RT-PCR Assays for SARS-CoV-2 Detection
Authors
KeywordsSARS-CoV-2
COVID-19
single-tube nested
RT-PCR
evaluation
Issue Date2020
PublisherMolecular Diversity Preservation International. The Journal's web site is located at http://www.mdpi.org/ijms
Citation
International Journal of Molecular Sciences, 2020, v. 21 n. 16, p. article no. 5674 How to Cite?
AbstractSensitive molecular assays are critical for coronavirus disease 2019 (COVID-19) diagnosis. Here, we designed and evaluated two single-tube nested (STN) real-time RT-PCR assays, targeting SARS-CoV-2 RdRp/Hel and N genes. Both STN assays had a low limit of detection and did not cross react with other human coronaviruses and respiratory viruses. Using 213 initial respiratory specimens from suspected COVID-19 patients, the sensitivity of both the STN COVID-19-RdRp/Hel and the STN COVID-19-N assays was 100% (99/99), while that of the comparator non-nested N assay was 95% (94/99). Among 108 follow-up specimens from confirmed COVID-19 patients who tested negative by the non-nested COVID-19-RdRp/Hel assay, 28 (25.9%) were positive for SARS-CoV-2 by the STN COVID-19-RdRp/Hel or the STN COVID-19-N assay. To evaluate the performance of our novel STN assays in pooled specimens, we created four sample pools, with each pool consisting of one low positive specimen and 49 negative specimens. While the non-nested COVID-19-RdRp/Hel assay was positive in only one of four sample pools (25%), both of the STN assays were positive in two of four samples pools (50%). In conclusion, the STN assays are highly sensitive and specific for SARS-CoV-2 detection. Their boosted sensitivity offers advantages in non-traditional COVID-19 testing algorithms such as saliva screening and pooled sample screening during massive screening.
Persistent Identifierhttp://hdl.handle.net/10722/304684
ISSN
2023 Impact Factor: 4.9
2023 SCImago Journal Rankings: 1.179
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorYip, CCY-
dc.contributor.authorSridhar, S-
dc.contributor.authorLeung, KH-
dc.contributor.authorNg, ACK-
dc.contributor.authorChan, KH-
dc.contributor.authorChan, JFW-
dc.contributor.authorTsang, OTY-
dc.contributor.authorHung, IFN-
dc.contributor.authorCheng, VCC-
dc.contributor.authorYuen, KY-
dc.contributor.authorTo, KKW-
dc.date.accessioned2021-10-05T02:33:40Z-
dc.date.available2021-10-05T02:33:40Z-
dc.date.issued2020-
dc.identifier.citationInternational Journal of Molecular Sciences, 2020, v. 21 n. 16, p. article no. 5674-
dc.identifier.issn1661-6596-
dc.identifier.urihttp://hdl.handle.net/10722/304684-
dc.description.abstractSensitive molecular assays are critical for coronavirus disease 2019 (COVID-19) diagnosis. Here, we designed and evaluated two single-tube nested (STN) real-time RT-PCR assays, targeting SARS-CoV-2 RdRp/Hel and N genes. Both STN assays had a low limit of detection and did not cross react with other human coronaviruses and respiratory viruses. Using 213 initial respiratory specimens from suspected COVID-19 patients, the sensitivity of both the STN COVID-19-RdRp/Hel and the STN COVID-19-N assays was 100% (99/99), while that of the comparator non-nested N assay was 95% (94/99). Among 108 follow-up specimens from confirmed COVID-19 patients who tested negative by the non-nested COVID-19-RdRp/Hel assay, 28 (25.9%) were positive for SARS-CoV-2 by the STN COVID-19-RdRp/Hel or the STN COVID-19-N assay. To evaluate the performance of our novel STN assays in pooled specimens, we created four sample pools, with each pool consisting of one low positive specimen and 49 negative specimens. While the non-nested COVID-19-RdRp/Hel assay was positive in only one of four sample pools (25%), both of the STN assays were positive in two of four samples pools (50%). In conclusion, the STN assays are highly sensitive and specific for SARS-CoV-2 detection. Their boosted sensitivity offers advantages in non-traditional COVID-19 testing algorithms such as saliva screening and pooled sample screening during massive screening.-
dc.languageeng-
dc.publisherMolecular Diversity Preservation International. The Journal's web site is located at http://www.mdpi.org/ijms-
dc.relation.ispartofInternational Journal of Molecular Sciences-
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.subjectSARS-CoV-2-
dc.subjectCOVID-19-
dc.subjectsingle-tube nested-
dc.subjectRT-PCR-
dc.subjectevaluation-
dc.titleDevelopment and Evaluation of Novel and Highly Sensitive Single-Tube Nested Real-Time RT-PCR Assays for SARS-CoV-2 Detection-
dc.typeArticle-
dc.identifier.emailYip, CCY: yipcyril@hku.hk-
dc.identifier.emailSridhar, S: sid8998@hku.hk-
dc.identifier.emailLeung, KH: khl17@hku.hk-
dc.identifier.emailChan, KH: chankh2@hkucc.hku.hk-
dc.identifier.emailChan, JFW: jfwchan@hku.hk-
dc.identifier.emailHung, IFN: ivanhung@hkucc.hku.hk-
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hk-
dc.identifier.emailTo, KKW: kelvinto@hku.hk-
dc.identifier.authorityYip, CCY=rp01721-
dc.identifier.authoritySridhar, S=rp02249-
dc.identifier.authorityChan, KH=rp01921-
dc.identifier.authorityChan, JFW=rp01736-
dc.identifier.authorityHung, IFN=rp00508-
dc.identifier.authorityYuen, KY=rp00366-
dc.identifier.authorityTo, KKW=rp01384-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.3390/ijms21165674-
dc.identifier.pmid32784770-
dc.identifier.pmcidPMC7461039-
dc.identifier.scopuseid_2-s2.0-85089407723-
dc.identifier.hkuros325985-
dc.identifier.volume21-
dc.identifier.issue16-
dc.identifier.spagearticle no. 5674-
dc.identifier.epagearticle no. 5674-
dc.identifier.isiWOS:000565064000001-
dc.publisher.placeSwitzerland-

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