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 : 000053 ( PascalFrancis/Corpus ); précédent : 000052; suivant : 000054Development 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 ; Wiroj Pongtanapisit ; Wantanich Piroj ; Chutatip Srichunrasmi ; Somying SeesuaiSource :
- Southeast Asian journal of tropical medicine and public health [ 0125-1562 ] ; 2004.
Descripteurs français
- Pascal (Inist)
English descriptors
- KwdEn :
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.
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NO : | PASCAL 04-0591783 INIST |
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ET : | 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 |
AU : | CHANTRATITA (Wasun); PONGTANAPISIT (Wiroj); PIROJ (Wantanich); SRICHUNRASMI (Chutatip); SEESUAI (Somying) |
AF : | Virology and Molecular Microbiology Unit, Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University/Bangkok/Thaïlande (1 aut., 2 aut., 3 aut., 4 aut., 5 aut.) |
DT : | Publication en série; Niveau analytique |
SO : | Southeast Asian journal of tropical medicine and public health; ISSN 0125-1562; Coden SJTMAK; Thaïlande; Da. 2004; Vol. 35; No. 3; Pp. 623-629; Bibl. 11 ref. |
LA : | Anglais |
EA : | 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. |
CC : | 002B01 |
FD : | Etude comparative; Réaction chaîne polymérase RT; Sonde; Syndrome respiratoire aigu sévère; Coronavirus; Médecine tropicale |
FG : | Virose; Infection; Coronaviridae; Nidovirales; Virus; Appareil respiratoire pathologie; Poumon pathologie |
ED : | Comparative study; Reverse transcription polymerase chain reaction; Probe; Severe acute respiratory syndrome; Coronavirus; Tropical medicine |
EG : | Viral disease; Infection; Coronaviridae; Nidovirales; Virus; Respiratory disease; Lung disease |
SD : | Estudio comparativo; Reacción cadena polimerasa transcripción inversa; Sonda; Síndrome respiratorio agudo severo; Coronavirus; Medicina tropical |
LO : | INIST-19778.354000122499500230 |
ID : | 04-0591783 |
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Pascal:04-0591783Le 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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<server><NO>PASCAL 04-0591783 INIST</NO>
<ET>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</ET>
<AU>CHANTRATITA (Wasun); PONGTANAPISIT (Wiroj); PIROJ (Wantanich); SRICHUNRASMI (Chutatip); SEESUAI (Somying)</AU>
<AF>Virology and Molecular Microbiology Unit, Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University/Bangkok/Thaïlande (1 aut., 2 aut., 3 aut., 4 aut., 5 aut.)</AF>
<DT>Publication en série; Niveau analytique</DT>
<SO>Southeast Asian journal of tropical medicine and public health; ISSN 0125-1562; Coden SJTMAK; Thaïlande; Da. 2004; Vol. 35; No. 3; Pp. 623-629; Bibl. 11 ref.</SO>
<LA>Anglais</LA>
<EA>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.</EA>
<CC>002B01</CC>
<FD>Etude comparative; Réaction chaîne polymérase RT; Sonde; Syndrome respiratoire aigu sévère; Coronavirus; Médecine tropicale</FD>
<FG>Virose; Infection; Coronaviridae; Nidovirales; Virus; Appareil respiratoire pathologie; Poumon pathologie</FG>
<ED>Comparative study; Reverse transcription polymerase chain reaction; Probe; Severe acute respiratory syndrome; Coronavirus; Tropical medicine</ED>
<EG>Viral disease; Infection; Coronaviridae; Nidovirales; Virus; Respiratory disease; Lung disease</EG>
<SD>Estudio comparativo; Reacción cadena polimerasa transcripción inversa; Sonda; Síndrome respiratorio agudo severo; Coronavirus; Medicina tropical</SD>
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