SrasV1, Main, Exploration, bibRecord, 005390

Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus

Identifieur interne : 005390 ( Main/Exploration ); précédent : 005389; suivant : 005391

Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus

Auteurs : Hong Thi Cam Thai ; Mai Quynh Le ; Cuong Duc Vuong ; Manmohan Parida ; Harumi Minekawa ; Tsugunori Notomi ; Futoshi Hasebe ; Kouichi Morita

Source :

Journal of Clinical Microbiology [ 0095-1137 ] ; 2004.

RBID : PMC:404656

Descripteurs français

KwdFr :
- ADN viral (génétique), Amorces ADN (génétique), Données de séquences moléculaires, Humains, Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Séquence nucléotidique, Techniques d'amplification d'acides nucléiques (), Virologie (), Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ADN viral, Amorces ADN, Virus du SRAS.
- isolement et purification : Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- Données de séquences moléculaires, Humains, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Séquence nucléotidique, Techniques d'amplification d'acides nucléiques, Virologie.

English descriptors

KwdEn :
- Base Sequence, DNA Primers (genetics), DNA, Viral (genetics), Humans, Molecular Sequence Data, Nucleic Acid Amplification Techniques (statistics & numerical data), Polymerase Chain Reaction (methods), Polymerase Chain Reaction (statistics & numerical data), SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Virology (methods), Virology (statistics & numerical data).
MESH :
- chemical , genetics : DNA Primers, DNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Polymerase Chain Reaction, Virology.
- statistics & numerical data : Nucleic Acid Amplification Techniques, Polymerase Chain Reaction, Virology.
- virology : Severe Acute Respiratory Syndrome.
- Base Sequence, Humans, Molecular Sequence Data, Sensitivity and Specificity.

Abstract

The development and evaluation of a one-step single-tube accelerated real-time quantitative reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assay is reported for rapid detection of the severe acute respiratory syndrome coronavirus (SARS-CoV) replicase gene. A total of 49 samples (15 throat washes, 13 throat swabs, and 21 combined throat and nasal swabs) collected from patients admitted to the Hanoi-French and Ninhbinh hospitals in Vietnam during the SARS epidemic were evaluated and compared to conventional RT-PCR. The RT-LAMP assay demonstrated 100-fold-greater sensitivity, with a detection limit of 0.01 PFU. The sensitivity and specificity of RT-LAMP assay for detecting viral RNA in clinical specimens with regard to RT-PCR were 100 and 87%, respectively. The specificity of the RT-LAMP assay was further validated by restriction analysis as well as nucleotide sequencing of the amplified product. The concentration of virus in most of the clinical samples was 0.1 PFU (0.1 to 10² PFU), as determined from the standard curve of SARS RT-LAMP and based on the time of positivity. The assay procedure is quite simple, wherein the amplification is carried out in a single tube under isothermal conditions at 63°C, and the result can be obtained in less than 1 h (as early as 11 min). Thus, the RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection and will be useful for rapid and reliable clinical diagnosis of SARS-CoV in developing countries.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC404656

DOI: 10.1128/JCM.42.5.1956-1961.2004
PubMed: 15131154
PubMed Central: 404656

Affiliations:

Le document en format XML

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<term>Humans</term>
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<term>Nucleic Acid Amplification Techniques (statistics & numerical data)</term>
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<front><div type="abstract" xml:lang="en"><p>The development and evaluation of a one-step single-tube accelerated real-time quantitative reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assay is reported for rapid detection of the severe acute respiratory syndrome coronavirus (SARS-CoV) replicase gene. A total of 49 samples (15 throat washes, 13 throat swabs, and 21 combined throat and nasal swabs) collected from patients admitted to the Hanoi-French and Ninhbinh hospitals in Vietnam during the SARS epidemic were evaluated and compared to conventional RT-PCR. The RT-LAMP assay demonstrated 100-fold-greater sensitivity, with a detection limit of 0.01 PFU. The sensitivity and specificity of RT-LAMP assay for detecting viral RNA in clinical specimens with regard to RT-PCR were 100 and 87%, respectively. The specificity of the RT-LAMP assay was further validated by restriction analysis as well as nucleotide sequencing of the amplified product. The concentration of virus in most of the clinical samples was 0.1 PFU (0.1 to 10<sup>2</sup>
 PFU), as determined from the standard curve of SARS RT-LAMP and based on the time of positivity. The assay procedure is quite simple, wherein the amplification is carried out in a single tube under isothermal conditions at 63°C, and the result can be obtained in less than 1 h (as early as 11 min). Thus, the RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection and will be useful for rapid and reliable clinical diagnosis of SARS-CoV in developing countries.</p>
</div>
</front>
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<affiliations><list></list>
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<name sortKey="Le, Mai Quynh" sort="Le, Mai Quynh" uniqKey="Le M" first="Mai Quynh" last="Le">Mai Quynh Le</name>
<name sortKey="Minekawa, Harumi" sort="Minekawa, Harumi" uniqKey="Minekawa H" first="Harumi" last="Minekawa">Harumi Minekawa</name>
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<name sortKey="Thai, Hong Thi Cam" sort="Thai, Hong Thi Cam" uniqKey="Thai H" first="Hong Thi Cam" last="Thai">Hong Thi Cam Thai</name>
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Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome Coronavirus

Development and Evaluation of a Novel Loop-Mediated Isothermal Amplification Method for Rapid Detection of Severe Acute Respiratory Syndrome 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

Pour générer des pages wiki

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