SrasV1, Main, Exploration, bibRecord, 004402

Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR

Identifieur interne : 004402 ( Main/Exploration ); précédent : 004401; suivant : 004403

Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR

Auteurs : Els Keyaerts [Belgique] ; Leen Vijgen [Belgique] ; Piet Maes [Belgique] ; Griet Duson [Belgique] ; Johan Neyts [Belgique] ; Marc Van Ranst [Belgique]

Source :

International journal of infectious diseases [ 1201-9712 ] ; 2006.

RBID : Pascal:06-0096173

Descripteurs français

Pascal (Inist)
- Syndrome respiratoire aigu sévère, Réaction chaîne polymérase RT, Réaction chaîne polymérase, Charge virale, Analyse quantitative, Norme, Coronavirus.
Wicri :
- topic : Analyse quantitative, Norme.

English descriptors

KwdEn :
- Coronavirus, Polymerase chain reaction, Quantitative analysis, Reverse transcription polymerase chain reaction, Severe acute respiratory syndrome, Standards, Viral load.

Abstract

Introduction: Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first occurred in humans in the People's Republic of China in November 2002 and has subsequently spread worldwide. A novel virus belonging to the Coronaviridae family has been identified as the cause of this pulmonary disease. The severity of the disease combined with its rapid spread requires the development of fast and sensitive diagnostic assays. Results: A real-time quantitative RT-PCR was designed in the nsp11 region of the replicase 1B domain of the SARS-coronavirus (SARS-CoV) genome. To evaluate this quantitative RT-PCR, cRNA standards were constructed by in vitro transcription of SARS-CoV Frankfurt 1 RNA using T7 RNA polymerase, followed by real-time RT-PCR. The assay allowed quantitation over a range of 10² to 10⁸ RNA copies per reaction. Conclusions: Extrapolated to clinical samples, this novel assay has a detection range of 10⁴ to 1010 copies of viral genome equivalents per millilitre. In comparison to the current de facto cRNA Artus Biotech standard, the in-house cRNA standard gives a 100-fold higher absolute quantity, suggesting a possible underestimation of the viral load when using the Artus Biotech standard.

Affiliations:

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Le document en format XML

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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus</term>
<term>Polymerase chain reaction</term>
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<front><div type="abstract" xml:lang="en">Introduction: Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first occurred in humans in the People's Republic of China in November 2002 and has subsequently spread worldwide. A novel virus belonging to the Coronaviridae family has been identified as the cause of this pulmonary disease. The severity of the disease combined with its rapid spread requires the development of fast and sensitive diagnostic assays. Results: A real-time quantitative RT-PCR was designed in the nsp11 region of the replicase 1B domain of the SARS-coronavirus (SARS-CoV) genome. To evaluate this quantitative RT-PCR, cRNA standards were constructed by in vitro transcription of SARS-CoV Frankfurt 1 RNA using T7 RNA polymerase, followed by real-time RT-PCR. The assay allowed quantitation over a range of 10<sup>2</sup>
 to 10<sup>8</sup>
 RNA copies per reaction. Conclusions: Extrapolated to clinical samples, this novel assay has a detection range of 10<sup>4</sup>
 to 1010 copies of viral genome equivalents per millilitre. In comparison to the current de facto cRNA Artus Biotech standard, the in-house cRNA standard gives a 100-fold higher absolute quantity, suggesting a possible underestimation of the viral load when using the Artus Biotech standard.</div>
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Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR

Viral load quantitation of SARS-coronavirus RNA using a one-step real-time RT-PCR

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri