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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

Identifieur interne : 000053 ( PascalFrancis/Corpus ); précédent : 000052; suivant : 000054

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 ; Wiroj Pongtanapisit ; Wantanich Piroj ; Chutatip Srichunrasmi ; Somying Seesuai

Source :

RBID : Pascal:04-0591783

Descripteurs français

English descriptors

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.

Notice en format standard (ISO 2709)

Pour connaître la documentation sur le format Inist Standard.

pA  
A01 01  1    @0 0125-1562
A02 01      @0 SJTMAK
A03   1    @0 Southeast asian j. trop. med. public health
A05       @2 35
A06       @2 3
A08 01  1  ENG  @1 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
A11 01  1    @1 CHANTRATITA (Wasun)
A11 02  1    @1 PONGTANAPISIT (Wiroj)
A11 03  1    @1 PIROJ (Wantanich)
A11 04  1    @1 SRICHUNRASMI (Chutatip)
A11 05  1    @1 SEESUAI (Somying)
A14 01      @1 Virology and Molecular Microbiology Unit, Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University @2 Bangkok @3 THA @Z 1 aut. @Z 2 aut. @Z 3 aut. @Z 4 aut. @Z 5 aut.
A20       @1 623-629
A21       @1 2004
A23 01      @0 ENG
A43 01      @1 INIST @2 19778 @5 354000122499500230
A44       @0 0000 @1 © 2004 INIST-CNRS. All rights reserved.
A45       @0 11 ref.
A47 01  1    @0 04-0591783
A60       @1 P
A61       @0 A
A64 01  1    @0 Southeast Asian journal of tropical medicine and public health
A66 01      @0 THA
C01 01    ENG  @0 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.
C02 01  X    @0 002B01
C03 01  X  FRE  @0 Etude comparative @5 02
C03 01  X  ENG  @0 Comparative study @5 02
C03 01  X  SPA  @0 Estudio comparativo @5 02
C03 02  X  FRE  @0 Réaction chaîne polymérase RT @5 03
C03 02  X  ENG  @0 Reverse transcription polymerase chain reaction @5 03
C03 02  X  SPA  @0 Reacción cadena polimerasa transcripción inversa @5 03
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C03 03  X  ENG  @0 Probe @5 05
C03 03  X  SPA  @0 Sonda @5 05
C03 04  X  FRE  @0 Syndrome respiratoire aigu sévère @2 NM @5 06
C03 04  X  ENG  @0 Severe acute respiratory syndrome @2 NM @5 06
C03 04  X  SPA  @0 Síndrome respiratorio agudo severo @2 NM @5 06
C03 05  X  FRE  @0 Coronavirus @2 NW @5 08
C03 05  X  ENG  @0 Coronavirus @2 NW @5 08
C03 05  X  SPA  @0 Coronavirus @2 NW @5 08
C03 06  X  FRE  @0 Médecine tropicale @5 09
C03 06  X  ENG  @0 Tropical medicine @5 09
C03 06  X  SPA  @0 Medicina tropical @5 09
C07 01  X  FRE  @0 Virose @2 NM
C07 01  X  ENG  @0 Viral disease @2 NM
C07 01  X  SPA  @0 Virosis @2 NM
C07 02  X  FRE  @0 Infection @2 NM
C07 02  X  ENG  @0 Infection @2 NM
C07 02  X  SPA  @0 Infección @2 NM
C07 03  X  FRE  @0 Coronaviridae @2 NW
C07 03  X  ENG  @0 Coronaviridae @2 NW
C07 03  X  SPA  @0 Coronaviridae @2 NW
C07 04  X  FRE  @0 Nidovirales @2 NW
C07 04  X  ENG  @0 Nidovirales @2 NW
C07 04  X  SPA  @0 Nidovirales @2 NW
C07 05  X  FRE  @0 Virus @2 NW
C07 05  X  ENG  @0 Virus @2 NW
C07 05  X  SPA  @0 Virus @2 NW
C07 06  X  FRE  @0 Appareil respiratoire pathologie @5 37
C07 06  X  ENG  @0 Respiratory disease @5 37
C07 06  X  SPA  @0 Aparato respiratorio patología @5 37
C07 07  X  FRE  @0 Poumon pathologie @5 38
C07 07  X  ENG  @0 Lung disease @5 38
C07 07  X  SPA  @0 Pulmón patología @5 38
N21       @1 341
N44 01      @1 OTO
N82       @1 OTO

Format Inist (serveur)

NO : PASCAL 04-0591783 INIST
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:04-0591783

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<fC03 i1="04" i2="X" l="ENG">
<s0>Severe acute respiratory syndrome</s0>
<s2>NM</s2>
<s5>06</s5>
</fC03>
<fC03 i1="04" i2="X" l="SPA">
<s0>Síndrome respiratorio agudo severo</s0>
<s2>NM</s2>
<s5>06</s5>
</fC03>
<fC03 i1="05" i2="X" l="FRE">
<s0>Coronavirus</s0>
<s2>NW</s2>
<s5>08</s5>
</fC03>
<fC03 i1="05" i2="X" l="ENG">
<s0>Coronavirus</s0>
<s2>NW</s2>
<s5>08</s5>
</fC03>
<fC03 i1="05" i2="X" l="SPA">
<s0>Coronavirus</s0>
<s2>NW</s2>
<s5>08</s5>
</fC03>
<fC03 i1="06" i2="X" l="FRE">
<s0>Médecine tropicale</s0>
<s5>09</s5>
</fC03>
<fC03 i1="06" i2="X" l="ENG">
<s0>Tropical medicine</s0>
<s5>09</s5>
</fC03>
<fC03 i1="06" i2="X" l="SPA">
<s0>Medicina tropical</s0>
<s5>09</s5>
</fC03>
<fC07 i1="01" i2="X" l="FRE">
<s0>Virose</s0>
<s2>NM</s2>
</fC07>
<fC07 i1="01" i2="X" l="ENG">
<s0>Viral disease</s0>
<s2>NM</s2>
</fC07>
<fC07 i1="01" i2="X" l="SPA">
<s0>Virosis</s0>
<s2>NM</s2>
</fC07>
<fC07 i1="02" i2="X" l="FRE">
<s0>Infection</s0>
<s2>NM</s2>
</fC07>
<fC07 i1="02" i2="X" l="ENG">
<s0>Infection</s0>
<s2>NM</s2>
</fC07>
<fC07 i1="02" i2="X" l="SPA">
<s0>Infección</s0>
<s2>NM</s2>
</fC07>
<fC07 i1="03" i2="X" l="FRE">
<s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="ENG">
<s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="SPA">
<s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="FRE">
<s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="ENG">
<s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="SPA">
<s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="05" i2="X" l="FRE">
<s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="05" i2="X" l="ENG">
<s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="05" i2="X" l="SPA">
<s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="06" i2="X" l="FRE">
<s0>Appareil respiratoire pathologie</s0>
<s5>37</s5>
</fC07>
<fC07 i1="06" i2="X" l="ENG">
<s0>Respiratory disease</s0>
<s5>37</s5>
</fC07>
<fC07 i1="06" i2="X" l="SPA">
<s0>Aparato respiratorio patología</s0>
<s5>37</s5>
</fC07>
<fC07 i1="07" i2="X" l="FRE">
<s0>Poumon pathologie</s0>
<s5>38</s5>
</fC07>
<fC07 i1="07" i2="X" l="ENG">
<s0>Lung disease</s0>
<s5>38</s5>
</fC07>
<fC07 i1="07" i2="X" l="SPA">
<s0>Pulmón patología</s0>
<s5>38</s5>
</fC07>
<fN21>
<s1>341</s1>
</fN21>
<fN44 i1="01">
<s1>OTO</s1>
</fN44>
<fN82>
<s1>OTO</s1>
</fN82>
</pA>
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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>
<LO>INIST-19778.354000122499500230</LO>
<ID>04-0591783</ID>
</server>
</inist>
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