Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus.
J Clin Microbiol
; 52(1): 67-75, 2014 Jan.
Article
en En
| MEDLINE
| ID: mdl-24153118
A new human coronavirus (CoV), subsequently named Middle East respiratory syndrome (MERS)-CoV, was first reported in Saudi Arabia in September 2012. In response, we developed two real-time reverse transcription-PCR (rRT-PCR) assays targeting the MERS-CoV nucleocapsid (N) gene and evaluated these assays as a panel with a previously published assay targeting the region upstream of the MERS-CoV envelope gene (upE) for the detection and confirmation of MERS-CoV infection. All assays detected ≤10 copies/reaction of quantified RNA transcripts, with a linear dynamic range of 8 log units and 1.3 × 10(-3) 50% tissue culture infective doses (TCID50)/ml of cultured MERS-CoV per reaction. All assays performed comparably with respiratory, serum, and stool specimens spiked with cultured virus. No false-positive amplifications were obtained with other human coronaviruses or common respiratory viral pathogens or with 336 diverse clinical specimens from non-MERS-CoV cases; specimens from two confirmed MERS-CoV cases were positive with all assay signatures. In June 2012, the U.S. Food and Drug Administration authorized emergency use of the rRT-PCR assay panel as an in vitro diagnostic test for MERS-CoV. A kit consisting of the three assay signatures and a positive control was assembled and distributed to public health laboratories in the United States and internationally to support MERS-CoV surveillance and public health responses.
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Infecciones del Sistema Respiratorio
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Infecciones por Coronavirus
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Coronavirus
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Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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Técnicas de Diagnóstico Molecular
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Reacción en Cadena en Tiempo Real de la Polimerasa
Tipo de estudio:
Diagnostic_studies
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Evaluation_studies
País/Región como asunto:
America do norte
Idioma:
En
Revista:
J Clin Microbiol
Año:
2014
Tipo del documento:
Article