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.
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.
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Galaxia
G
Cortés-Hinojosa
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Frances M D
FM
Gulland
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Tracey
T
Goldstein
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Stephanie
S
Venn-Watson
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Rebecca
R
Rivera
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Linda L
LL
Archer
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Thomas B
TB
Waltzek
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
Gregory C
GC
Gray
Departments of Small Animal Clinical Sciences (Cortes-Hinojosa, Archer, Wellehan) and Infectious Diseases and Pathology (Waltzek), College of Veterinary Medicine, University of Florida, Gainesville, FL.|Global Health Institute, Duke University, Durham, NC (Gray).|The Marine Mammal Center, Sausalito, CA (Gulland).|Wildlife Health Center, School of Veterinary Medicine, University of California-Davis, Davis, CA (Goldstein).|National Marine Mammal Foundation, San Diego, CA (Venn-Watson).|Hubbs-SeaWorld Research Institute, San Diego, CA (Rivera).
James F X
JF
Wellehan
D000256:Adenoviridae / Q000235:genetics / Q000302:isolation & purification*; D000257:Adenoviridae Infections / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D000818:Animals; D000835:Animals, Wild; D002140:California / Q000453:epidemiology; D015995:Prevalence; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards / Q000662:veterinary*; D046070:Sea Lions
Adenovirus; California sea lions; pinnipeds; quantitative PCR