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PLoS One ; 4(9): e7081, 2009 Sep 17.
Artículo en Inglés | MEDLINE | ID: mdl-19759905

RESUMEN

In 2007, the Centers for Disease Control and Prevention (CDC) reported that Human adenovirus type 14 (HAdV-14) infected 106 military personnel and was responsible for the death of one U.S. soldier at Lackland Air Force Base in Texas. Identification of the responsible adenovirus, which had not previously been seen in North America and for which rapid diagnostic tools were unavailable, required retrospective analysis at reference laboratories. Initial quarantine measures were also reliant on relatively slow traditional PCR analysis at other locations. To address this problem, we developed a real-time PCR assay that detects a 225 base pair sequence in the HAdV-14a hexon gene. Fifty-one oropharyngeal swab specimens from the Naval Health Research Center, San Diego, CA and Advanced Diagnostic Laboratory, Lackland AFB, TX were used to validate the new assay. The described assay detected eight of eight and 19 of 19 confirmed HAdV-14a clinical isolates in two separate cohorts from respiratory disease outbreaks. The real-time PCR assay had a wide dynamic range, detecting from 10(2) to 10(7) copies of genomic DNA per reaction. The assay did not cross-react with other adenoviruses, influenza, respiratory syncytial virus, or common respiratory tract bacteria. The described assay is easy to use, sensitive and specific for HAdV-14a in clinical throat swab specimens, and very rapid since turnaround time is less than four hours to obtain an answer.


Asunto(s)
Infecciones por Adenovirus Humanos/diagnóstico , Infecciones por Adenovirus Humanos/virología , Adenovirus Humanos/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Secuencia de Bases , Estudios de Cohortes , Brotes de Enfermedades , Humanos , Datos de Secuencia Molecular , ARN Viral/genética , Reproducibilidad de los Resultados , Enfermedades Respiratorias/diagnóstico , Enfermedades Respiratorias/virología , Sensibilidad y Especificidad , Homología de Secuencia de Ácido Nucleico , Factores de Tiempo
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