File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Evaluation of simple nucleic acid extraction methods for the detection of SARS-CoV-2 in nasopharyngeal and saliva specimens during global shortage of extraction kits

TitleEvaluation of simple nucleic acid extraction methods for the detection of SARS-CoV-2 in nasopharyngeal and saliva specimens during global shortage of extraction kits
Authors
KeywordsSARS-CoV-2
Proteinase K
Detection
RT-PCR
COVID-19
Issue Date2020
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jcv
Citation
Journal of Clinical Virology, 2020, v. 129, p. article no. 104519 How to Cite?
AbstractBackground: The severe shortage of nucleic acid extraction kits during the current COVID-19 pandemic represents a key limiting factor in testing capacity. Objectives: This study compared the results of SARS-CoV-2 RT-PCR using different simple nucleic acid extraction methods on nasopharyngeal and saliva specimens. Study design: Fifty nasopharyngeal swab and saliva specimens previously tested positive for SARS-CoV-2 were retrieved. Three different methods of nucleic acid extraction were compared. The first method involves incubating the specimen with proteinase K, and then heat treatment at 98 °C for 5 min (PKH); the second method involves heat treatment at 98 °C for 5 min without proteinase K pre-incubation (heat only); the third method involves no pre-processing steps (direct). The products from all 3 methods were tested by SARS-CoV-2 RT-PCR. Results: PKH had significantly higher positive rate in SARS-CoV-2 RT-PCR (80 %) than those of heat only (58 %; P = 0.001) or direct (56 %; P = 0.002). The median Ct value was significantly earlier for PKH (median Ct: 37.0, IQR 31.7–40) than that of heat only (median Ct: 40, IQR 36.2–41; P < 0.0001) and direct (median Ct, 37.5; IQR 33.9–41.0; P = 0.0049). Subgroup analysis showed that PKH had higher detection rate, lower limit of detection and earlier Ct values than the other two groups for both NPS and saliva specimens. Conclusions: PKH pre-processing resulted in the highest detection rate of SARS-CoV-2 by RT-PCR, and represents an alternative method for nucleic acid extraction when commercial extraction kits are not available.
Persistent Identifierhttp://hdl.handle.net/10722/285299
ISSN
2019 Impact Factor: 2.777
2015 SCImago Journal Rankings: 1.503
PubMed Central ID

 

DC FieldValueLanguage
dc.contributor.authorChu, AWH-
dc.contributor.authorChan, WM-
dc.contributor.authorIp, JD-
dc.contributor.authorYip, CCY-
dc.contributor.authorChan, JFW-
dc.contributor.authorYuen, KY-
dc.contributor.authorTo, KKW-
dc.date.accessioned2020-08-18T03:52:10Z-
dc.date.available2020-08-18T03:52:10Z-
dc.date.issued2020-
dc.identifier.citationJournal of Clinical Virology, 2020, v. 129, p. article no. 104519-
dc.identifier.issn1386-6532-
dc.identifier.urihttp://hdl.handle.net/10722/285299-
dc.description.abstractBackground: The severe shortage of nucleic acid extraction kits during the current COVID-19 pandemic represents a key limiting factor in testing capacity. Objectives: This study compared the results of SARS-CoV-2 RT-PCR using different simple nucleic acid extraction methods on nasopharyngeal and saliva specimens. Study design: Fifty nasopharyngeal swab and saliva specimens previously tested positive for SARS-CoV-2 were retrieved. Three different methods of nucleic acid extraction were compared. The first method involves incubating the specimen with proteinase K, and then heat treatment at 98 °C for 5 min (PKH); the second method involves heat treatment at 98 °C for 5 min without proteinase K pre-incubation (heat only); the third method involves no pre-processing steps (direct). The products from all 3 methods were tested by SARS-CoV-2 RT-PCR. Results: PKH had significantly higher positive rate in SARS-CoV-2 RT-PCR (80 %) than those of heat only (58 %; P = 0.001) or direct (56 %; P = 0.002). The median Ct value was significantly earlier for PKH (median Ct: 37.0, IQR 31.7–40) than that of heat only (median Ct: 40, IQR 36.2–41; P < 0.0001) and direct (median Ct, 37.5; IQR 33.9–41.0; P = 0.0049). Subgroup analysis showed that PKH had higher detection rate, lower limit of detection and earlier Ct values than the other two groups for both NPS and saliva specimens. Conclusions: PKH pre-processing resulted in the highest detection rate of SARS-CoV-2 by RT-PCR, and represents an alternative method for nucleic acid extraction when commercial extraction kits are not available.-
dc.languageeng-
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jcv-
dc.relation.ispartofJournal of Clinical Virology-
dc.subjectSARS-CoV-2-
dc.subjectProteinase K-
dc.subjectDetection-
dc.subjectRT-PCR-
dc.subjectCOVID-19-
dc.titleEvaluation of simple nucleic acid extraction methods for the detection of SARS-CoV-2 in nasopharyngeal and saliva specimens during global shortage of extraction kits-
dc.typeArticle-
dc.identifier.emailChu, AWH: awhchu@hku.hk-
dc.identifier.emailChan, WM: mbally@hku.hk-
dc.identifier.emailIp, JD: jdip1007@connect.hku.hk-
dc.identifier.emailYip, CCY: yipcyril@hku.hk-
dc.identifier.emailChan, JFW: jfwchan@hku.hk-
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hk-
dc.identifier.emailTo, KKW: kelvinto@hku.hk-
dc.identifier.authorityYip, CCY=rp01721-
dc.identifier.authorityChan, JFW=rp01736-
dc.identifier.authorityYuen, KY=rp00366-
dc.identifier.authorityTo, KKW=rp01384-
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1016/j.jcv.2020.104519-
dc.identifier.pmid32629187-
dc.identifier.pmcidPMC7309780-
dc.identifier.scopuseid_2-s2.0-85087309967-
dc.identifier.hkuros312813-
dc.identifier.volume129-
dc.identifier.spagearticle no. 104519-
dc.identifier.epagearticle no. 104519-
dc.publisher.placeNetherlands-

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats