Development and validation of a quantitative PCR for rapid and specific detection of California sea lion adenovirus 1 and prevalence in wild and managed populations

• Published in: Journal of veterinary diagnostic investigation Vol. 29; no. 2; p. 193
• Main Authors: Cortés-Hinojosa, Galaxia, Gulland, Frances M D, Goldstein, Tracey, Venn-Watson, Stephanie, Rivera, Rebecca, Archer, Linda L, Waltzek, Thomas B, Gray, Gregory C, Wellehan, Jr, James F X
• Format: Journal Article
• Language: English
• Published: United States 01.03.2017
• Subjects: Adenoviridae - genetics; Adenoviridae - isolation & purification; Adenoviridae Infections - epidemiology; Adenoviridae Infections - veterinary; Adenoviridae Infections - virology; Animals; Animals, Wild; California - epidemiology; Prevalence; Real-Time Polymerase Chain Reaction - standards; Real-Time Polymerase Chain Reaction - veterinary; Sea Lions; California sea lions; Adenovirus; quantitative PCR; pinnipeds
• ISSN: 1943-4936
• DOI: 10.1177/1040638716689113

California sea lion adenovirus 1 (CSLAdV-1) has been associated with hepatitis and enteritis in several wild and captive populations of diverse pinniped species. Currently available tests have been limited to pan-adenoviral polymerase chain reaction (PCR) followed by sequencing. We present the development of a quantitative probe-hybridization PCR (qPCR) assay for rapid, sensitive, and specific detection of this virus in California sea lions ( Zalophus californianus) and other pinnipeds. This assay did not amplify other mammalian adenoviruses and is able to detect consistently down to 10 viral copies per well. Compared with the gold standard conventional pan-adenovirus PCR/sequencing assay, diagnostic sensitivity and specificity of 100% and 88.2% were found, respectively. The lower diagnostic specificity of this qPCR assay may be the result of the lower limit of detection of this assay compared with the gold standard rather than the result of detection of true false-positives.