Application of cross-priming amplification (CPA) for detection of fowl adenovirus (FAdV) strains

• Published in: Archives of virology 2015-04, Vol.160 (4), p.1005-1013
• Main Authors: Niczyporuk, Jowita Samanta, Woźniakowski, Grzegorz, Samorek-Salamonowicz, Elżbieta
• Format: Article
• Language: English
• Subjects: Adenoviridae Infections - diagnosis; Adenoviridae Infections - veterinary; Adenoviridae Infections - virology; Adenovirus; Adenoviruses; Anemia; Animals; Antibodies; Atadenovirus; Aviadenovirus; Aviadenovirus - classification; Aviadenovirus - genetics; Aviadenovirus - isolation & purification; Aviadenovirus - physiology; Base Sequence; Biomedical and Life Sciences; Biomedicine; Birds; Chick Embryo; Chickens; DNA Primers - genetics; Embryos; Hepatitis; Infections; Infectious Diseases; Infectious laryngotracheitis virus; Marek's disease herpesvirus; Medical Microbiology; Molecular Sequence Data; Nucleic Acid Amplification Techniques - methods; Original; Original Article; Pathogens; Phylogeny; Poultry; Poultry Diseases - diagnosis; Poultry Diseases - virology; Virology; Virulence
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/s00705-015-2355-9

Fowl adenoviruses (FAdVs) are widely distributed among chickens. Detection of FAdVs is mainly accomplished by virus isolation, serological assays, various polymerase chain reaction (PCR) assays, and loop-mediated isothermal amplification (LAMP). To increase the diagnostic capacity of currently applied techniques, cross-priming amplification (CPA) for the detection of the FAdV hexon gene was developed. The single CPA assay was optimised to detect all serotypes 1-8a-8b-11 representing the species Fowl aviadenovirus A-E . The optimal temperature and incubation time were determined to be 68 °C for 2 h. Using different incubation temperatures, it was possible to differentiate some FAdV serotypes. The results were recorded after addition of SYBR Green I ® dye, which produced a greenish fluorescence under UV light. The CPA products separated by gel electrophoresis showed different “ladder-like” patterns for the different serotypes. The assay was specific for all serotypes of FAdV, and no cross-reactivity was observed with members of the genus Atadenovirus , duck atadenovirus A (egg drop syndrome virus EDS-76 [EDSV]) or control samples containing Marek’s disease virus (MDV), infectious laryngotracheitis virus (ILTV) or chicken anaemia virus (CAV). The results of the newly developed FAdV-CPA were compared with those of real-time PCR. The sensitivity of CPA was equal to that of real-time PCR and reached 10 −2.0 TCID 50 , but the CPA method was more rapid and cheaper than the PCR systems. CPA is a highly specific, sensitive, efficient, and rapid tool for detection of all FAdV serotypes. This is the first report on the application of CPA for detection of FAdV strains.