SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses.
Identifieur interne : 001638 ( Ncbi/Checkpoint ); précédent : 001637; suivant : 001639SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses.
Auteurs : S. Escutenaire [Suède] ; N. Mohamed ; M. Isaksson ; P. Thorén ; B. Klingeborn ; S. Belák ; M. Berg ; J. BlombergSource :
- Archives of virology [ 0304-8608 ] ; 2007.
Descripteurs français
- KwdFr :
- ADN viral (génétique), ARN complémentaire (génétique), ARN viral (génétique), Amorces ADN (génétique), Animaux, Colorants fluorescents, Composés chimiques organiques, Coronaviridae (), Coronaviridae (génétique), Coronaviridae (isolement et purification), Données de séquences moléculaires, Humains, Phylogénie, RT-PCR (), Reproductibilité des résultats, Sensibilité et spécificité, Séquence nucléotidique.
- MESH :
- génétique : ADN viral, ARN complémentaire, ARN viral, Amorces ADN, Coronaviridae.
- isolement et purification : Coronaviridae.
- Animaux, Colorants fluorescents, Composés chimiques organiques, Coronaviridae, Données de séquences moléculaires, Humains, Phylogénie, RT-PCR, Reproductibilité des résultats, Sensibilité et spécificité, Séquence nucléotidique.
English descriptors
- KwdEn :
- Animals, Base Sequence, Coronaviridae (classification), Coronaviridae (genetics), Coronaviridae (isolation & purification), DNA Primers (genetics), DNA, Viral (genetics), Fluorescent Dyes, Humans, Molecular Sequence Data, Organic Chemicals, Phylogeny, RNA, Complementary (genetics), RNA, Viral (genetics), Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction (methods), Reverse Transcriptase Polymerase Chain Reaction (statistics & numerical data), Sensitivity and Specificity.
- MESH :
- chemical , genetics : DNA Primers, DNA, Viral, RNA, Complementary, RNA, Viral.
- classification : Coronaviridae.
- genetics : Coronaviridae.
- isolation & purification : Coronaviridae.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- statistics & numerical data : Reverse Transcriptase Polymerase Chain Reaction.
- Animals, Base Sequence, Fluorescent Dyes, Humans, Molecular Sequence Data, Organic Chemicals, Phylogeny, Reproducibility of Results, Sensitivity and Specificity.
Abstract
Coronaviruses are etiologic agents of respiratory and enteric diseases in humans and in animals. In this study, a one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on SYBR Green chemistry and degenerate primers was developed for the generic detection of coronaviruses. The primers, designed in the open reading frame 1b, enabled the detection of 32 animal coronaviruses including strains of canine coronavirus, feline coronavirus, transmissible gastroenteritis virus (TGEV), bovine coronavirus (BCoV), murine hepatitis virus (MHV) and infectious bronchitis virus (IBV). A specific amplification was also observed with the human coronaviruses (HCoV) HCoV-NL63, HCoV-OC43, HCoV-229E and severe acute respiratory syndrome coronavirus (SARS-CoV). The real-time RT-PCR detected down to 10 cRNA copies from TGEV, BCoV, SARS-CoV and IBV. In addition, the assay exhibited a high sensitivity and specificity on clinical samples from different animal species. The developed assay represents a potential tool for laboratory diagnostics and for detecting still uncharacterized coronaviruses.
DOI: 10.1007/s00705-006-0840-x
PubMed: 16941059
Affiliations:
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pubmed:16941059Le document en format XML
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In this study, a one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on SYBR Green chemistry and degenerate primers was developed for the generic detection of coronaviruses. The primers, designed in the open reading frame 1b, enabled the detection of 32 animal coronaviruses including strains of canine coronavirus, feline coronavirus, transmissible gastroenteritis virus (TGEV), bovine coronavirus (BCoV), murine hepatitis virus (MHV) and infectious bronchitis virus (IBV). A specific amplification was also observed with the human coronaviruses (HCoV) HCoV-NL63, HCoV-OC43, HCoV-229E and severe acute respiratory syndrome coronavirus (SARS-CoV). The real-time RT-PCR detected down to 10 cRNA copies from TGEV, BCoV, SARS-CoV and IBV. In addition, the assay exhibited a high sensitivity and specificity on clinical samples from different animal species. The developed assay represents a potential tool for laboratory diagnostics and for detecting still uncharacterized coronaviruses.</div></front></TEI><affiliations><list><country><li>Suède</li></country></list><tree><noCountry><name sortKey="Belak, S" sort="Belak, S" uniqKey="Belak S" first="S" last="Belák">S. Belák</name><name sortKey="Berg, M" sort="Berg, M" uniqKey="Berg M" first="M" last="Berg">M. Berg</name><name sortKey="Blomberg, J" sort="Blomberg, J" uniqKey="Blomberg J" first="J" last="Blomberg">J. Blomberg</name><name sortKey="Isaksson, M" sort="Isaksson, M" uniqKey="Isaksson M" first="M" last="Isaksson">M. Isaksson</name><name sortKey="Klingeborn, B" sort="Klingeborn, B" uniqKey="Klingeborn B" first="B" last="Klingeborn">B. Klingeborn</name><name sortKey="Mohamed, N" sort="Mohamed, N" uniqKey="Mohamed N" first="N" last="Mohamed">N. Mohamed</name><name sortKey="Thoren, P" sort="Thoren, P" uniqKey="Thoren P" first="P" last="Thorén">P. Thorén</name></noCountry><country name="Suède"><noRegion><name sortKey="Escutenaire, S" sort="Escutenaire, S" uniqKey="Escutenaire S" first="S" last="Escutenaire">S. Escutenaire</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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