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Selected Publication:

Publication type: Journal Article
Document type: Full Paper

Year: 2012

Author(s): Günes, A; Marek, A; Grafl, B; Berger, E; Hess, M

Title: Real-time PCR assay for universal detection and quantitation of all five species of fowl adenoviruses (FAdV-A to FAdV-E).

Source: J Virol Methods. 2012; 183(2):147-153

Authors Vetmeduni Vienna:

Grafl Beatrice
Hess Michael
Marek Ana

Vetmed Research Units
University Clinic for Poultry and Fish Medicine, Clinical Unit of Poultry Medicine

The present study describes the development of a SYBR Green based real-time polymerase chain reaction (real-time PCR) for detection and quantitation of all fowl adenovirus (FAdV) species. Primers were designed based on conserved nucleotide sequences within the 52K gene. Ten-fold serial dilutions of a vector DNA were used as standard for quantitation. The real-time PCR had an efficiency of 98%, a regression squared value of 0.999 and showed a range of 6.73-6.73×10(8) copies of FAdV DNA per reaction. The assay was highly specific for FAdVs and an exact quantitation of all 5 FAdV species (FAdV-A to FAdV-E) could be demonstrated. It was shown, that twelve FAdV serotypes (FAdV-1 to 8a, and 8b to 11) were detectable and quantifiable. Other viral genomes as well as uninfected chicken embryo liver (CEL) cells did not produce positive signal. Cloacal swabs were taken during the animal experiment, which was performed with all FAdV species. Shedding of FAdVs was investigated in cell culture, by conventional PCR and by the developed real-time PCR. The real-time PCR was found more sensitive than cell culture and conventional PCR. Detection and quantitation of FAdVs in different type of samples was possible by the new real-time PCR.

Keywords Pubmed: Adenoviridae Infections/diagnosis
Adenoviridae Infections/immunology
Adenoviridae Infections/veterinary*
Adenoviridae Infections/virology
Antibodies, Viral/blood
Cell Line
Chick Embryo
Fowl adenovirus A/genetics*
Fowl adenovirus A/immunology
Fowl adenovirus A/physiology
Genome, Viral
Molecular Diagnostic Techniques
Molecular Typing*
Poultry Diseases/diagnosis*
Poultry Diseases/immunology
Poultry Diseases/virology
Real-Time Polymerase Chain Reaction*/standards
Reference Standards
Sensitivity and Specificity
Transition Temperature
Virus Shedding

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