Evaluation and validation of a real-time PCR assay for detection and quantitation of human adenovirus 14 from clinical samples

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 previo...

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Bibliographic Details
Published in:	PloS one 2009-09, Vol.4 (9), p.e7081-e7081
Main Authors:	Metzgar, David, Skochko, Greg, Gibbins, Carl, Hudson, Nolan, Lott, Lisa, Jones, Morris S
Format:	Article
Language:	eng
Subjects:	Adenovirus Infections, Human - diagnosis Adenovirus Infections, Human - virology Adenoviruses Adenoviruses, Human - genetics Air bases Armed forces Assaying Bacteria Base Sequence Clinical isolates Cohort Studies Deoxyribonucleic acid Diagnostic software Diagnostic systems Disease control Disease Outbreaks DNA Epidemics Genomes Humans Identification Influenza Laboratories Medical research Military personnel Molecular Biology Molecular Sequence Data Occupational exposure Outbreaks Pharynx Polymerase chain reaction Public Health and Epidemiology/Screening Quantitation Real time Reproducibility of Results Respiratory diseases Respiratory syncytial virus Respiratory tract Respiratory tract diseases Respiratory Tract Diseases - diagnosis Respiratory Tract Diseases - virology Reverse Transcriptase Polymerase Chain Reaction - methods RNA, Viral - genetics Sensitivity and Specificity Sequence Homology, Nucleic Acid Time Factors Vibrio vulnificus Viral proteins Virology/Diagnosis Viruses
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Summary:	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.
ISSN:	1932-6203 1932-6203