Automated extraction protocol for quantification of SARS-coronavirus RNA in serum: an evaluation study.
Identifieur interne : 002352 ( PubMed/Curation ); précédent : 002351; suivant : 002353Automated extraction protocol for quantification of SARS-coronavirus RNA in serum: an evaluation study.
Auteurs : Rossa W K. Chiu [Hong Kong] ; Yongjie Jin ; Grace T Y. Chung ; Wing-Bong Lui ; Anthony T C. Chan ; Wilina Lim ; Y M Dennis LoSource :
- BMC infectious diseases [ 1471-2334 ] ; 2006.
Descripteurs français
- KwdFr :
- ARN viral (isolement et purification), ARN viral (sang), Analyse de régression, Charge virale, Humains, RT-PCR, Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Techniques de diagnostic moléculaire (), Trousses de réactifs pour diagnostic, Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Virus du SRAS.
- isolement et purification : ARN viral, Virus du SRAS.
- sang : ARN viral.
- virologie : Syndrome respiratoire aigu sévère.
- Analyse de régression, Charge virale, Humains, RT-PCR, Sensibilité et spécificité, Techniques de diagnostic moléculaire, Trousses de réactifs pour diagnostic.
English descriptors
- KwdEn :
- Humans, Molecular Diagnostic Techniques (methods), RNA, Viral (blood), RNA, Viral (isolation & purification), Reagent Kits, Diagnostic, Regression Analysis, Reverse Transcriptase Polymerase Chain Reaction, SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Viral Load.
- MESH :
- chemical , blood : RNA, Viral.
- chemical , isolation & purification : RNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Molecular Diagnostic Techniques.
- virology : Severe Acute Respiratory Syndrome.
- Humans, Reagent Kits, Diagnostic, Regression Analysis, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity, Viral Load.
Abstract
We have previously developed a test for the diagnosis and prognostic assessment of the severe acute respiratory syndrome (SARS) based on the detection of the SARS-coronavirus RNA in serum by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). In this study, we evaluated the feasibility of automating the serum RNA extraction procedure in order to increase the throughput of the assay.
DOI: 10.1186/1471-2334-6-20
PubMed: 16466582
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pubmed:16466582Le document en format XML
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<author><name sortKey="Chiu, Rossa W K" sort="Chiu, Rossa W K" uniqKey="Chiu R" first="Rossa W K" last="Chiu">Rossa W K. Chiu</name>
<affiliation wicri:level="1"><nlm:affiliation>Centre for Emerging Infectious Diseases, The Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong. rossachiu@cuhk.edu.hk</nlm:affiliation>
<country xml:lang="fr">Hong Kong</country>
<wicri:regionArea>Centre for Emerging Infectious Diseases, The Chinese University of Hong Kong, Prince of Wales Hospital</wicri:regionArea>
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<author><name sortKey="Jin, Yongjie" sort="Jin, Yongjie" uniqKey="Jin Y" first="Yongjie" last="Jin">Yongjie Jin</name>
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<author><name sortKey="Chung, Grace T Y" sort="Chung, Grace T Y" uniqKey="Chung G" first="Grace T Y" last="Chung">Grace T Y. Chung</name>
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<author><name sortKey="Lui, Wing Bong" sort="Lui, Wing Bong" uniqKey="Lui W" first="Wing-Bong" last="Lui">Wing-Bong Lui</name>
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<author><name sortKey="Chan, Anthony T C" sort="Chan, Anthony T C" uniqKey="Chan A" first="Anthony T C" last="Chan">Anthony T C. Chan</name>
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<author><name sortKey="Lim, Wilina" sort="Lim, Wilina" uniqKey="Lim W" first="Wilina" last="Lim">Wilina Lim</name>
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<term>Molecular Diagnostic Techniques (methods)</term>
<term>RNA, Viral (blood)</term>
<term>RNA, Viral (isolation & purification)</term>
<term>Reagent Kits, Diagnostic</term>
<term>Regression Analysis</term>
<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (isolation & purification)</term>
<term>Sensitivity and Specificity</term>
<term>Severe Acute Respiratory Syndrome (diagnosis)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Viral Load</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (isolement et purification)</term>
<term>ARN viral (sang)</term>
<term>Analyse de régression</term>
<term>Charge virale</term>
<term>Humains</term>
<term>RT-PCR</term>
<term>Sensibilité et spécificité</term>
<term>Syndrome respiratoire aigu sévère (diagnostic)</term>
<term>Syndrome respiratoire aigu sévère (virologie)</term>
<term>Techniques de diagnostic moléculaire ()</term>
<term>Trousses de réactifs pour diagnostic</term>
<term>Virus du SRAS (génétique)</term>
<term>Virus du SRAS (isolement et purification)</term>
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<keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>RNA, Viral</term>
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<keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>RNA, Viral</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>ARN viral</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Molecular Diagnostic Techniques</term>
</keywords>
<keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>ARN viral</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Humans</term>
<term>Reagent Kits, Diagnostic</term>
<term>Regression Analysis</term>
<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>Sensitivity and Specificity</term>
<term>Viral Load</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Analyse de régression</term>
<term>Charge virale</term>
<term>Humains</term>
<term>RT-PCR</term>
<term>Sensibilité et spécificité</term>
<term>Techniques de diagnostic moléculaire</term>
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<front><div type="abstract" xml:lang="en">We have previously developed a test for the diagnosis and prognostic assessment of the severe acute respiratory syndrome (SARS) based on the detection of the SARS-coronavirus RNA in serum by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). In this study, we evaluated the feasibility of automating the serum RNA extraction procedure in order to increase the throughput of the assay.</div>
</front>
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<Month>04</Month>
<Day>01</Day>
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<DateRevised><Year>2019</Year>
<Month>12</Month>
<Day>10</Day>
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<Month>Feb</Month>
<Day>09</Day>
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<Title>BMC infectious diseases</Title>
<ISOAbbreviation>BMC Infect. Dis.</ISOAbbreviation>
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<ArticleTitle>Automated extraction protocol for quantification of SARS-coronavirus RNA in serum: an evaluation study.</ArticleTitle>
<Pagination><MedlinePgn>20</MedlinePgn>
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<Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">We have previously developed a test for the diagnosis and prognostic assessment of the severe acute respiratory syndrome (SARS) based on the detection of the SARS-coronavirus RNA in serum by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). In this study, we evaluated the feasibility of automating the serum RNA extraction procedure in order to increase the throughput of the assay.</AbstractText>
<AbstractText Label="METHODS" NlmCategory="METHODS">An automated nucleic acid extraction platform using the MagNA Pure LC instrument (Roche Diagnostics) was evaluated. We developed a modified protocol in compliance with the recommended biosafety guidelines from the World Health Organization based on the use of the MagNA Pure total nucleic acid large volume isolation kit for the extraction of SARS-coronavirus RNA. The modified protocol was compared with a column-based extraction kit (QIAamp viral RNA mini kit, Qiagen) for quantitative performance, analytical sensitivity and precision.</AbstractText>
<AbstractText Label="RESULTS" NlmCategory="RESULTS">The newly developed automated protocol was shown to be free from carry-over contamination and have comparable performance with other standard protocols and kits designed for the MagNA Pure LC instrument. However, the automated method was found to be less sensitive, less precise and led to consistently lower serum SARS-coronavirus concentrations when compared with the column-based extraction method.</AbstractText>
<AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">As the diagnostic efficiency and prognostic value of the serum SARS-CoV RNA RT-PCR test is critically associated with the analytical sensitivity and quantitative performance contributed both by the RNA extraction and RT-PCR components of the test, we recommend the use of the column-based manual RNA extraction method.</AbstractText>
</Abstract>
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<ForeName>Rossa W K</ForeName>
<Initials>RW</Initials>
<AffiliationInfo><Affiliation>Centre for Emerging Infectious Diseases, The Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong. rossachiu@cuhk.edu.hk</Affiliation>
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