Preclinical Evaluation of Two Real-Time, Reverse Transcription-PCR Assays for Detection of the Severe Acute Respiratory Syndrome Coronavirus
Identifieur interne : 000641 ( Ncbi/Checkpoint ); précédent : 000640; suivant : 000642Preclinical Evaluation of Two Real-Time, Reverse Transcription-PCR Assays for Detection of the Severe Acute Respiratory Syndrome Coronavirus
Auteurs : Adam M. Bressler [Géorgie (pays)] ; Frederick S. Nolte [Géorgie (pays)]Source :
- Journal of Clinical Microbiology [ 0095-1137 ] ; 2004.
Descripteurs français
- KwdFr :
- ARN viral (génétique), ARN viral (isolement et purification), Analyse de régression, Cadres ouverts de lecture (génétique), Fèces (virologie), Humains, Liquide de lavage bronchoalvéolaire (virologie), Microbiologie de l'eau, RT-PCR (), Reproductibilité des résultats, Syndrome respiratoire aigu sévère (diagnostic), Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ARN viral, Cadres ouverts de lecture, Virus du SRAS.
- isolement et purification : ARN viral, Virus du SRAS.
- virologie : Fèces, Liquide de lavage bronchoalvéolaire.
- Analyse de régression, Humains, Microbiologie de l'eau, RT-PCR, Reproductibilité des résultats.
English descriptors
- KwdEn :
- Bronchoalveolar Lavage Fluid (virology), Feces (virology), Humans, Open Reading Frames (genetics), RNA, Viral (genetics), RNA, Viral (isolation & purification), Regression Analysis, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction (methods), SARS Virus (genetics), SARS Virus (isolation & purification), Severe Acute Respiratory Syndrome (diagnosis), Water Microbiology.
- MESH :
- chemical , genetics : RNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : Open Reading Frames, SARS Virus.
- chemical , isolation & purification : RNA, Viral, SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- virology : Bronchoalveolar Lavage Fluid, Feces.
- Humans, Regression Analysis, Reproducibility of Results, Water Microbiology.
Abstract
We verified the analytical performance characteristics of a previously described real-time reverse transcription-PCR (RT-PCR) assay targeting the open reading frame (ORF) 1b region of the severe acute respiratory syndrome coronavirus (SARS-CoV) with RNA transcripts. We then compared it to a novel nucleocapsid gene real-time RT-PCR assay with genomic RNA. The assays differed only in the primer and probe sequences and final concentrations. A commercially available armored RNA (Ambion, Austin, Tex.) was evaluated as positive control for the ORF 1b assay. The analytical sensitivity, reproducibility, amplification efficiency, and dynamic range of the assays were similar. Both were specific for SARS-CoV as determined by testing against human CoV 229E and OC43, specimens from patients without SARS, and by BLAST searches of GenBank for primer and probe sequence homology. The armored RNA was found to be a suitable positive control for the ORF 1b assay that could be reliably recovered and amplified from a variety of clinical specimens.
Url:
DOI: 10.1128/JCM.42.3.987-991.2004
PubMed: 15004042
PubMed Central: 356893
Affiliations:
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PMC:356893Le document en format XML
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