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- Publisher Website: 10.1016/j.jcv.2003.08.004
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- PMID: 14522060
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Article: Early diagnosis of SARS Coronavirus infection by real time RT-PCR
Title | Early diagnosis of SARS Coronavirus infection by real time RT-PCR |
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Authors | |
Keywords | Early diagnosis Real time RT-PCR SARS Coronavirus |
Issue Date | 2003 |
Publisher | Elsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jcv |
Citation | Journal Of Clinical Virology, 2003, v. 28 n. 3, p. 233-238 How to Cite? |
Abstract | Background: A novel coronavirus was recently identified as the aetiological agent of Severe Acute Respiratory Syndrome (SARS). Molecular assays currently available for detection of SARS-coronavirus (SARS-Cov) have low sensitivity during the early stage of the illness. Objective: To develop and evaluate a sensitive diagnostic test for SARS by optimizing the viral RNA extraction methods and by applying real-time quantitative RT-PCR technology. Study design: 50 nasopharyngeal aspirate (NPA) samples collected from days 1-3 of disease onset from SARS patients in whom SARS CoV infections was subsequently serologically confirmed and 30 negative control samples were studied. Samples were tested by: (1) our first generation conventional RT-PCR assay with a routine RNA extraction method (Lancet 361 (2003) 1319), (2) our first generation conventional RT-PCR assay with a modified RNA extraction method, (3) a real-time quantitative RT-PCR assay with a modified RNA extraction method. Results: Of 50 NPA specimens collected during the first 3 days of illness, 11 (22%) were positive in our first generation RT-PCR assay. With a modification in the RNA extraction protocol, 22 (44%) samples were positive in the conventional RT-PCR assay. By combining the modified RNA extraction method and real-time quantitative PCR technology, 40 (80%) of these samples were positive in the real-time RT-PCR assay. No positive signal was observed in the negative controls. Conclusion: By optimizing RNA extraction methods and applying quantitative real time RT-PCR technologies, the sensitivity of tests for early diagnosis of SARS can be greatly enhanced. © 2003 Published by Elsevier B.V. |
Persistent Identifier | http://hdl.handle.net/10722/78825 |
ISSN | 2023 Impact Factor: 4.0 2023 SCImago Journal Rankings: 1.344 |
ISI Accession Number ID | |
References |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Poon, LLM | en_HK |
dc.contributor.author | Chan, KH | en_HK |
dc.contributor.author | Wong, OK | en_HK |
dc.contributor.author | Yam, WC | en_HK |
dc.contributor.author | Yuen, KY | en_HK |
dc.contributor.author | Guan, Y | en_HK |
dc.contributor.author | Lo, YMD | en_HK |
dc.contributor.author | Peiris, JSM | en_HK |
dc.date.accessioned | 2010-09-06T07:47:16Z | - |
dc.date.available | 2010-09-06T07:47:16Z | - |
dc.date.issued | 2003 | en_HK |
dc.identifier.citation | Journal Of Clinical Virology, 2003, v. 28 n. 3, p. 233-238 | en_HK |
dc.identifier.issn | 1386-6532 | en_HK |
dc.identifier.uri | http://hdl.handle.net/10722/78825 | - |
dc.description.abstract | Background: A novel coronavirus was recently identified as the aetiological agent of Severe Acute Respiratory Syndrome (SARS). Molecular assays currently available for detection of SARS-coronavirus (SARS-Cov) have low sensitivity during the early stage of the illness. Objective: To develop and evaluate a sensitive diagnostic test for SARS by optimizing the viral RNA extraction methods and by applying real-time quantitative RT-PCR technology. Study design: 50 nasopharyngeal aspirate (NPA) samples collected from days 1-3 of disease onset from SARS patients in whom SARS CoV infections was subsequently serologically confirmed and 30 negative control samples were studied. Samples were tested by: (1) our first generation conventional RT-PCR assay with a routine RNA extraction method (Lancet 361 (2003) 1319), (2) our first generation conventional RT-PCR assay with a modified RNA extraction method, (3) a real-time quantitative RT-PCR assay with a modified RNA extraction method. Results: Of 50 NPA specimens collected during the first 3 days of illness, 11 (22%) were positive in our first generation RT-PCR assay. With a modification in the RNA extraction protocol, 22 (44%) samples were positive in the conventional RT-PCR assay. By combining the modified RNA extraction method and real-time quantitative PCR technology, 40 (80%) of these samples were positive in the real-time RT-PCR assay. No positive signal was observed in the negative controls. Conclusion: By optimizing RNA extraction methods and applying quantitative real time RT-PCR technologies, the sensitivity of tests for early diagnosis of SARS can be greatly enhanced. © 2003 Published by Elsevier B.V. | en_HK |
dc.language | eng | en_HK |
dc.publisher | Elsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jcv | en_HK |
dc.relation.ispartof | Journal of Clinical Virology | en_HK |
dc.rights | Journal of Clinical Virology. Copyright © Elsevier BV. | en_HK |
dc.subject | Early diagnosis | en_HK |
dc.subject | Real time RT-PCR | en_HK |
dc.subject | SARS Coronavirus | en_HK |
dc.subject.mesh | Humans | en_HK |
dc.subject.mesh | Nasopharynx - virology | en_HK |
dc.subject.mesh | RNA, Viral - analysis - isolation & purification | en_HK |
dc.subject.mesh | Reverse Transcriptase Polymerase Chain Reaction - methods | en_HK |
dc.subject.mesh | SARS Virus - isolation & purification | en_HK |
dc.subject.mesh | Sensitivity and Specificity | en_HK |
dc.subject.mesh | Severe Acute Respiratory Syndrome - diagnosis - virology | en_HK |
dc.subject.mesh | Time Factors | en_HK |
dc.title | Early diagnosis of SARS Coronavirus infection by real time RT-PCR | en_HK |
dc.type | Article | en_HK |
dc.identifier.openurl | http://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1386-6532&volume=28&issue=3&spage=233&epage=8&date=2003&atitle=Early+diagnosis+of+SARS+Coronavirus+infection+by+real+time+RT-PCR | en_HK |
dc.identifier.email | Poon, LLM: llmpoon@hkucc.hku.hk | en_HK |
dc.identifier.email | Yam, WC: wcyam@hkucc.hku.hk | en_HK |
dc.identifier.email | Yuen, KY: kyyuen@hkucc.hku.hk | en_HK |
dc.identifier.email | Guan, Y: yguan@hkucc.hku.hk | en_HK |
dc.identifier.email | Peiris, JSM: malik@hkucc.hku.hk | en_HK |
dc.identifier.authority | Poon, LLM=rp00484 | en_HK |
dc.identifier.authority | Yam, WC=rp00313 | en_HK |
dc.identifier.authority | Yuen, KY=rp00366 | en_HK |
dc.identifier.authority | Guan, Y=rp00397 | en_HK |
dc.identifier.authority | Peiris, JSM=rp00410 | en_HK |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1016/j.jcv.2003.08.004 | en_HK |
dc.identifier.pmid | 14522060 | - |
dc.identifier.scopus | eid_2-s2.0-0141763635 | en_HK |
dc.identifier.hkuros | 87556 | en_HK |
dc.relation.references | http://www.scopus.com/mlt/select.url?eid=2-s2.0-0141763635&selection=ref&src=s&origin=recordpage | en_HK |
dc.identifier.volume | 28 | en_HK |
dc.identifier.issue | 3 | en_HK |
dc.identifier.spage | 233 | en_HK |
dc.identifier.epage | 238 | en_HK |
dc.identifier.isi | WOS:000187448800002 | - |
dc.publisher.place | Netherlands | en_HK |
dc.identifier.scopusauthorid | Poon, LLM=7005441747 | en_HK |
dc.identifier.scopusauthorid | Chan, KH=7406034307 | en_HK |
dc.identifier.scopusauthorid | Wong, OK=7004813969 | en_HK |
dc.identifier.scopusauthorid | Yam, WC=7004281720 | en_HK |
dc.identifier.scopusauthorid | Yuen, KY=36078079100 | en_HK |
dc.identifier.scopusauthorid | Guan, Y=7202924055 | en_HK |
dc.identifier.scopusauthorid | Lo, YMD=7401935391 | en_HK |
dc.identifier.scopusauthorid | Peiris, JSM=7005486823 | en_HK |
dc.identifier.issnl | 1386-6532 | - |