CovidV1, Main, Exploration, bibRecord, 001076

Development and comparison of the real-time amplification based methods: NASBA-Beacon, RT-PCR Taqman and RT-PCR hybridization probe assays: For the qualitative detection of SARS coronavirus

Identifieur interne : 001076 ( Main/Exploration ); précédent : 001075; suivant : 001077

Development and comparison of the real-time amplification based methods: NASBA-Beacon, RT-PCR Taqman and RT-PCR hybridization probe assays: For the qualitative detection of SARS coronavirus

Auteurs : Wasun Chantratita [Thaïlande] ; Wiroj Pongtanapisit [Thaïlande] ; Wantanich Piroj [Thaïlande] ; Chutatip Srichunrasmi [Thaïlande] ; Somying Seesuai [Thaïlande]

Source :

Southeast Asian journal of tropical medicine and public health [ 0125-1562 ] ; 2004.

RBID : Pascal:04-0591783

Descripteurs français

Pascal (Inist)
- Etude comparative, Réaction chaîne polymérase RT, Sonde, Syndrome respiratoire aigu sévère, Coronavirus, Médecine tropicale.

English descriptors

KwdEn :
- Comparative study, Coronavirus, Probe, Reverse transcription polymerase chain reaction, Severe acute respiratory syndrome, Tropical medicine.

Abstract

The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe based on Taqman (RT-PCR TaqMan assay), hybridization probe (RT-PCR hybridization probe assay) and a real-time assay with virus-specific molecular beacon probes (NASBA-Beacon assay). The analytical sensitivity or reproducibility of the test results among those three assays was compared. All assays yielded results by detecting SARS coronavirus targeting the BNI-1 region in less than 2 hours. RNA detection by all the formats was unaffected by the presence of human sputum. The limits of detection were at least 10 copies of input RNA for both RT-PCR formats (RT-PCR TaqMan and RT-PCR hybridization probe assays), while the NASBA-Beacon assay could detect as little as 1 copy per reaction, with high reproducibility of the coefficient of variation (CV) of <10. These results demonstrate that real-time NASBA provides a rapid and sensitive alternative to RT-PCR for the routine qualitative assay of sputum for SARS corona viral RNA detection.

Affiliations:

Thaïlande

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

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to stream PascalFrancis, to step Checkpoint: 000053
to stream Main, to step Merge: 001087
to stream Main, to step Curation: 001076

Le document en format XML

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<front><div type="abstract" xml:lang="en">The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe based on Taqman (RT-PCR TaqMan assay), hybridization probe (RT-PCR hybridization probe assay) and a real-time assay with virus-specific molecular beacon probes (NASBA-Beacon assay). The analytical sensitivity or reproducibility of the test results among those three assays was compared. All assays yielded results by detecting SARS coronavirus targeting the BNI-1 region in less than 2 hours. RNA detection by all the formats was unaffected by the presence of human sputum. The limits of detection were at least 10 copies of input RNA for both RT-PCR formats (RT-PCR TaqMan and RT-PCR hybridization probe assays), while the NASBA-Beacon assay could detect as little as 1 copy per reaction, with high reproducibility of the coefficient of variation (CV) of <10. These results demonstrate that real-time NASBA provides a rapid and sensitive alternative to RT-PCR for the routine qualitative assay of sputum for SARS corona viral RNA detection.</div>
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Development and comparison of the real-time amplification based methods: NASBA-Beacon, RT-PCR Taqman and RT-PCR hybridization probe assays: For the qualitative detection of SARS coronavirus

Development and comparison of the real-time amplification based methods: NASBA-Beacon, RT-PCR Taqman and RT-PCR hybridization probe assays: For the qualitative detection of SARS coronavirus

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