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Article: Clinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods

TitleClinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periods
Authors
KeywordsRapid diagnosis
Real-time PCR
SARS coronavirus
Issue Date2005
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jcv
Citation
Journal Of Clinical Virology, 2005, v. 33 n. 1, p. 19-24 How to Cite?
AbstractBackground: The protocols of WHO network laboratories facilitated development of rapid diagnosis for SARS coronavirus (CoV) using reverse transcription (RT)-PCR assays. However, several reports have shown that conventional and real-time PCR assays were very specific for SARS CoV but lack sensitivity depending on the assay, specimen, and time course of disease. Objective: To evaluate an automatic nucleic acid extraction system and two standardized real-time PCR assays for rapid diagnosis of SARS CoV during outbreak and post-epidemic periods in Hong Kong. Study design: Specimens from clinically suspected SARS patients collected during outbreak and post-epidemic periods were tested by an automatic nucleic acid extraction system followed by our first generation conventional RT-PCR and two standardized real-time PCR assays (Artus GmbH, Germany and Roche Diagnostics, Germany). Paired serum samples were assayed for increasing titer against SARS CoV. Results: In the SARS epidemic, Artus and Roche PCR assays exhibited sensitivities of 87% and 85% for respiratory specimens (n = 64), 91% and 88% for stool (n = 44), and 82% for urine (n = 29). A specificity of 100% was exhibited by both PCR assays except Artus attained only a 92% specificity for stool. For post-epidemic period, no SARS CoV was identified among 56 respiratory specimens by all PCR assays. Inhibitors to PCR assays were detected at an average rate of 7-8% among 202 clinical specimens. Conclusion: This study highlights the high throughput and performance of automatic RNA extraction in coordination with standardized real-time PCR assays suitable for large-scale routine diagnosis in case of future SARS epidemic. As no SARS CoV was detected among specimens collected during post-epidemic period, the positive predictive value of real-time PCR assays for detection of SARS CoV during low epidemic requires further evaluation. © 2004 Elsevier B.V. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/78974
ISSN
2015 Impact Factor: 2.647
2015 SCImago Journal Rankings: 1.503
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorYam, WCen_HK
dc.contributor.authorChan, KHen_HK
dc.contributor.authorChow, KHen_HK
dc.contributor.authorPoon, LLMen_HK
dc.contributor.authorLam, HYen_HK
dc.contributor.authorYuen, KYen_HK
dc.contributor.authorSeto, WHen_HK
dc.contributor.authorPeiris, JSMen_HK
dc.date.accessioned2010-09-06T07:49:03Z-
dc.date.available2010-09-06T07:49:03Z-
dc.date.issued2005en_HK
dc.identifier.citationJournal Of Clinical Virology, 2005, v. 33 n. 1, p. 19-24en_HK
dc.identifier.issn1386-6532en_HK
dc.identifier.urihttp://hdl.handle.net/10722/78974-
dc.description.abstractBackground: The protocols of WHO network laboratories facilitated development of rapid diagnosis for SARS coronavirus (CoV) using reverse transcription (RT)-PCR assays. However, several reports have shown that conventional and real-time PCR assays were very specific for SARS CoV but lack sensitivity depending on the assay, specimen, and time course of disease. Objective: To evaluate an automatic nucleic acid extraction system and two standardized real-time PCR assays for rapid diagnosis of SARS CoV during outbreak and post-epidemic periods in Hong Kong. Study design: Specimens from clinically suspected SARS patients collected during outbreak and post-epidemic periods were tested by an automatic nucleic acid extraction system followed by our first generation conventional RT-PCR and two standardized real-time PCR assays (Artus GmbH, Germany and Roche Diagnostics, Germany). Paired serum samples were assayed for increasing titer against SARS CoV. Results: In the SARS epidemic, Artus and Roche PCR assays exhibited sensitivities of 87% and 85% for respiratory specimens (n = 64), 91% and 88% for stool (n = 44), and 82% for urine (n = 29). A specificity of 100% was exhibited by both PCR assays except Artus attained only a 92% specificity for stool. For post-epidemic period, no SARS CoV was identified among 56 respiratory specimens by all PCR assays. Inhibitors to PCR assays were detected at an average rate of 7-8% among 202 clinical specimens. Conclusion: This study highlights the high throughput and performance of automatic RNA extraction in coordination with standardized real-time PCR assays suitable for large-scale routine diagnosis in case of future SARS epidemic. As no SARS CoV was detected among specimens collected during post-epidemic period, the positive predictive value of real-time PCR assays for detection of SARS CoV during low epidemic requires further evaluation. © 2004 Elsevier B.V. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jcven_HK
dc.relation.ispartofJournal of Clinical Virologyen_HK
dc.rightsJournal of Clinical Virology. Copyright © Elsevier BV.en_HK
dc.subjectRapid diagnosisen_HK
dc.subjectReal-time PCRen_HK
dc.subjectSARS coronavirusen_HK
dc.subject.meshDisease Outbreaksen_HK
dc.subject.meshHumansen_HK
dc.subject.meshPolymerase Chain Reaction - methodsen_HK
dc.subject.meshReagent Kits, Diagnosticen_HK
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_HK
dc.subject.meshSARS Virus - genetics - isolation & purificationen_HK
dc.subject.meshSensitivity and Specificityen_HK
dc.subject.meshSevere Acute Respiratory Syndrome - diagnosis - epidemiology - virologyen_HK
dc.subject.meshTime Factorsen_HK
dc.titleClinical evaluation of real-time PCR assays for rapid diagnosis of SARS coronavirus during outbreak and post-epidemic periodsen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1386-6532&volume=33&issue=1&spage=19&epage=24&date=2005&atitle=Clinical+evaluation+of+real-time+PCR+assays+for+rapid+diagnosis+of+SARS+coronavirus+during+outbreak+and+post-epidemic+periods.en_HK
dc.identifier.emailYam, WC: wcyam@hkucc.hku.hken_HK
dc.identifier.emailChow, KH: khchowb@hku.hken_HK
dc.identifier.emailPoon, LLM: llmpoon@hkucc.hku.hken_HK
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hken_HK
dc.identifier.emailPeiris, JSM: malik@hkucc.hku.hken_HK
dc.identifier.authorityYam, WC=rp00313en_HK
dc.identifier.authorityChow, KH=rp00370en_HK
dc.identifier.authorityPoon, LLM=rp00484en_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.identifier.authorityPeiris, JSM=rp00410en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.jcv.2004.09.029en_HK
dc.identifier.pmid15797361-
dc.identifier.scopuseid_2-s2.0-15944406346en_HK
dc.identifier.hkuros103926en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-15944406346&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume33en_HK
dc.identifier.issue1en_HK
dc.identifier.spage19en_HK
dc.identifier.epage24en_HK
dc.identifier.isiWOS:000229030500004-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridYam, WC=7004281720en_HK
dc.identifier.scopusauthoridChan, KH=35338760600en_HK
dc.identifier.scopusauthoridChow, KH=7202180736en_HK
dc.identifier.scopusauthoridPoon, LLM=7005441747en_HK
dc.identifier.scopusauthoridLam, HY=35097472400en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK
dc.identifier.scopusauthoridSeto, WH=7005799377en_HK
dc.identifier.scopusauthoridPeiris, JSM=7005486823en_HK

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