High Influenza A Virus Infection Rates in Mallards Bred for Hunting in the Camargue, South of France

Vittecoq, Marion; Grandhomme, Viviane; Champagnon, Jocelyn; Guillemain, Matthieu; Crescenzo-Chaigne, Bernadette; Renaud, François; Thomas, Frédéric; Gauthier-Clerc, Michel; van der Werf, Sylvie

For 10 RT-PCR positive samples, 200 ml of specimen was inoculated in the allantoic cavity of 10-days-old embryonated hen's eggs. The allantoic fluid was harvested after 72 h at 37uC and influenza A virus was detected by hemagglutination assay with hen erythrocytes. Viral RNA was purified from allantoic fluid using a QIAamp viral RNA mini kit (Qiagen) following manufacturer's instructions. HA subtype of virus isolates was determined by RT-PCR and sequencing of the HA 0 cleavage site region using a universal set of primers as described previously [57] . HA sequence was analyzed with the basic local alignment search tool available from NCBI (data not shown) and confirmed by sequencing of the whole HA gene using a set of H10-specific primers (primer sequences available upon request). The NA subtype was deduced from this analysis and confirmed by RT-PCR and sequencing of 172 nt of the NA using a set of H10specific primers (primer sequences available upon request).