Evaluation and Validation of Real-Time PCR Assay Using J.B.A.I.D.S. System for Detection and Quantitation of Human Adenovirus 14 from Nasal Washes

Session III – Jones, Morris – Young Investigator Applicant

Title of Contribution: Evaluation and Validation of Real-Time PCR Assay Using J.B.A.I.D.S. System for Detection and Quantitation of Human Adenovirus 14 from Nasal Washes
Author(s): Morris S. Jones II1, Carl Gibbins1, N. Ryan Hudson1, David Metzgar2 and Lisa Lott3
Affiliation(s): 1David Grant Medical Center/ 2Naval Health Research Center/ 3Advanced Diagnostic Laboratory

Abstract: In 2007, the Centers for Disease Control and Prevention (CDC) reported that HAdV-14 infected 106 military personnel and was responsible for the death of one U.S. soldier at Lackland Air Force Base in Texas. To address this problem, we developed a real-time J.B.A.I.D.S. PCR assay that detects a 225 base pair sequence in the HAdV-B14 hexon gene. Twenty-four nasal wash 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 four of four and 21 of 21 confirmed HAdV-B14 clinical isolates in two separate cohorts from respiratory disease outbreaks. The lower limit of detection for our real-time J.B.A.I.D.S. PCR assay was calculated to be 10 genomic equivalents per PCR reaction. The assay did not cross-react with other adenoviruses, influenza, respiratory syncytial virus, or respiratory disease causing bacteria. Because of the ease of use, sensitivity, specificity, and frequency in HAdV-B14 infections in the U.S. military, we believe this assay has the potential to be useful as a clinical diagnostic tool for HAdV-B14 infection.