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Article: Detection of SARS Coronavirus in Patients with Severe Acute Respiratory Syndrome by Conventional and Real-Time Quantitative Reverse Transcription-PCR Assays

TitleDetection of SARS Coronavirus in Patients with Severe Acute Respiratory Syndrome by Conventional and Real-Time Quantitative Reverse Transcription-PCR Assays
Authors
Issue Date2004
PublisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.org
Citation
Clinical Chemistry, 2004, v. 50 n. 1, p. 67-72 How to Cite?
AbstractBackground: A novel coronavirus (CoV) was recently identified as the agent for severe acute respiratory syndrome (SARS). We compared the abilities of conventional and real-time reverse transcription-PCR (RT-PCR) assays to detect SARS CoV in clinical specimens. Methods: RNA samples isolated from nasopharyngeal aspirate (NPA; n = 170) and stool (n = 44) were reverse-transcribed and tested by our in-house conventional RT-PCR assay. We selected 98 NPA and 37 stool samples collected at different times after the onset of disease and tested them in a real-time quantitative RT-PCR specific for the open reading frame (ORF) 1b region of SARS CoV. Detection rates for the conventional and real-time quantitative RT-PCR assays were compared. To investigate the nature of viral RNA molecules in these clinical samples, we determined copy numbers of ORF 1b and nucleocapsid (N) gene sequences of SARS CoV. Results: The quantitative real-time RT-PCR assay was more sensitive than the conventional RT-PCR assay for detecting SARS CoV in samples collected early in the course of the disease. Real-time assays targeted at the ORF 1b region and the N gene revealed that copy numbers of ORF 1b and N gene sequences in clinical samples were similar. Conclusions: NPA and stool samples can be used for early diagnosis of SARS. The real-time quantitative RT-PCR assay for SARS CoV is potentially useful for early detection of SARS CoV. Our results suggest that genomic RNA is the predominant viral RNA species in clinical samples. © 2004 American Association for Clinical Chemistry.
Persistent Identifierhttp://hdl.handle.net/10722/79262
ISSN
2023 Impact Factor: 7.1
2023 SCImago Journal Rankings: 1.460
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorPoon, LLMen_HK
dc.contributor.authorChan, KHen_HK
dc.contributor.authorWong, OKen_HK
dc.contributor.authorCheung, TKWen_HK
dc.contributor.authorNg, Ien_HK
dc.contributor.authorZheng, Ben_HK
dc.contributor.authorSeto, WHen_HK
dc.contributor.authorYuen, KYen_HK
dc.contributor.authorGuan, Yen_HK
dc.contributor.authorPeiris, JSMen_HK
dc.date.accessioned2010-09-06T07:52:33Z-
dc.date.available2010-09-06T07:52:33Z-
dc.date.issued2004en_HK
dc.identifier.citationClinical Chemistry, 2004, v. 50 n. 1, p. 67-72en_HK
dc.identifier.issn0009-9147en_HK
dc.identifier.urihttp://hdl.handle.net/10722/79262-
dc.description.abstractBackground: A novel coronavirus (CoV) was recently identified as the agent for severe acute respiratory syndrome (SARS). We compared the abilities of conventional and real-time reverse transcription-PCR (RT-PCR) assays to detect SARS CoV in clinical specimens. Methods: RNA samples isolated from nasopharyngeal aspirate (NPA; n = 170) and stool (n = 44) were reverse-transcribed and tested by our in-house conventional RT-PCR assay. We selected 98 NPA and 37 stool samples collected at different times after the onset of disease and tested them in a real-time quantitative RT-PCR specific for the open reading frame (ORF) 1b region of SARS CoV. Detection rates for the conventional and real-time quantitative RT-PCR assays were compared. To investigate the nature of viral RNA molecules in these clinical samples, we determined copy numbers of ORF 1b and nucleocapsid (N) gene sequences of SARS CoV. Results: The quantitative real-time RT-PCR assay was more sensitive than the conventional RT-PCR assay for detecting SARS CoV in samples collected early in the course of the disease. Real-time assays targeted at the ORF 1b region and the N gene revealed that copy numbers of ORF 1b and N gene sequences in clinical samples were similar. Conclusions: NPA and stool samples can be used for early diagnosis of SARS. The real-time quantitative RT-PCR assay for SARS CoV is potentially useful for early detection of SARS CoV. Our results suggest that genomic RNA is the predominant viral RNA species in clinical samples. © 2004 American Association for Clinical Chemistry.en_HK
dc.languageengen_HK
dc.publisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.orgen_HK
dc.relation.ispartofClinical Chemistryen_HK
dc.subject.meshFeces - virologyen_HK
dc.subject.meshHumansen_HK
dc.subject.meshNasal Cavity - virologyen_HK
dc.subject.meshPharynx - virologyen_HK
dc.subject.meshRNA, Viral - analysisen_HK
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_HK
dc.subject.meshSARS Virus - genetics - isolation & purificationen_HK
dc.subject.meshSevere Acute Respiratory Syndrome - virologyen_HK
dc.subject.meshSpecimen Handlingen_HK
dc.titleDetection of SARS Coronavirus in Patients with Severe Acute Respiratory Syndrome by Conventional and Real-Time Quantitative Reverse Transcription-PCR Assaysen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0009-9147&volume=50&issue=1&spage=67&epage=72&date=2004&atitle=Detection+of+SARS+coronavirus+in+patients+with+severe+acute+respiratory+syndrome+by+conventional+and+real-time+quantitative+reverse+transcription-PCR+assays.en_HK
dc.identifier.emailPoon, LLM: llmpoon@hkucc.hku.hken_HK
dc.identifier.emailZheng, B: bzheng@hkucc.hku.hken_HK
dc.identifier.emailYuen, KY: kyyuen@hkucc.hku.hken_HK
dc.identifier.emailGuan, Y: yguan@hkucc.hku.hken_HK
dc.identifier.emailPeiris, JSM: malik@hkucc.hku.hken_HK
dc.identifier.authorityPoon, LLM=rp00484en_HK
dc.identifier.authorityZheng, B=rp00353en_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.identifier.authorityGuan, Y=rp00397en_HK
dc.identifier.authorityPeiris, JSM=rp00410en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1373/clinchem.2003.023663en_HK
dc.identifier.pmid14709637-
dc.identifier.scopuseid_2-s2.0-9144230030en_HK
dc.identifier.hkuros87587en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-9144230030&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume50en_HK
dc.identifier.issue1en_HK
dc.identifier.spage67en_HK
dc.identifier.epage72en_HK
dc.identifier.isiWOS:000188216900005-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridPoon, LLM=7005441747en_HK
dc.identifier.scopusauthoridChan, KH=7406034307en_HK
dc.identifier.scopusauthoridWong, OK=7004813969en_HK
dc.identifier.scopusauthoridCheung, TKW=16229531100en_HK
dc.identifier.scopusauthoridNg, I=8671050800en_HK
dc.identifier.scopusauthoridZheng, B=7201780588en_HK
dc.identifier.scopusauthoridSeto, WH=7005799377en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK
dc.identifier.scopusauthoridGuan, Y=7202924055en_HK
dc.identifier.scopusauthoridPeiris, JSM=7005486823en_HK
dc.identifier.issnl0009-9147-

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