The University of Georgia College of Veterinary Medicine

Georgia Veterinary Scholar Program

Georgia Veterinary Scholar	Faculty Mentor

April Chambers University of Georgia Class of 2007	Dr. David Suarez

Development of a Real-Time Reverse Transcriptase PCR Assay for Avian Influenza H2 Hemagglutin Subtypes

Avian influenza virus (AIV) is type A influenza. The natural reservoir of the virus is considered to be within the free flying bird population. Previously, the H2 hemagglutinin (HA) subtypes have been associated with low path avian influenza in poultry. H2 HA has historically been isolated from people, although has not been detected within the past twenty years. During an outbreak of avian influenza in commercial poultry, subtyping becomes important due to trade regulations as well as animal health. An RRT-PCR assay for H2 was developed and optimized using hydrolysis type probes. The gold standard for virus detection is virus isolation (VI) in embryonating chicken eggs followed by hemagglutinin and neuraminidase subtyping via serology with results taking 1-2 weeks compared to less than one day with RRT-PCR. Limit of detection was determined by comparisons with VI in embryonating chicken eggs. Specificity was determined with eight North American origin H2 wild bird isolates and a panel comprised of all 15 HA subtypes. The RRT-PCR detected seven out of the eight isolates and cross reacted with four other HA subtypes.