Polymerase chain reaction combined with restriction enzyme analysis for detection and differentiation of fowl adenoviruses

A polymerase chain reaction combined with restriction enzyme analysis was d eveloped for detection and differentiation of all 12 fowl adenovirus (FAdV) serotypes representing the five fowl adenovirus (A to E) species. For prim er design, the published sequences of the hexon proteins of FAdV1, FAdV8 an d FAdV9 were aligned and conserved regions in the two pedestal regions adja cent to the L1 loop region were determined. A primer pair ( hexon A/hexon B ) was constructed and was shown to amplify approximately 900 bp of the hexo n gene including the L1 loop region. An amplification product was detected using supernatant of infected cell cultures from all FAdV1 to FAdV12 refere nce strains used in our study. The sequence and the restriction patterns of the hexon A/B fragments of the 12 FAdV strains were determined and compare d. The successive use of four different endonucleases allowed the complete differentiation of the reference FAdV strains. Twenty-six fowl adenoviruses isolated during our routine virological diagnosis activities could all be amplified using hexon A/hexon B primers. Restriction analysis results showe d that 8/26 adenovirus strains contained two different FAdV types. FAdV4, F AdV12, FAdV1, FAdV5 and FAdV6 were the most frequently isolated.