Rapid Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H9 and H5
Abstract: A method of one-step reverse transcription polymerase chain reaction was developed to subtype hemagglutinin(HA) gene of Avian influenza virus(AIV).One pair of primers were designed on the basis of HA gene of AIV,allowing simultaneous detection of AIV subtype H_9 and H_5,and the specific amplicons were 579bp and 177bp in size respectively.The primers were specific for AIV(H_9 and H_5 subtype) with no cross-reaction to RNA from the chicken muscle and other chicken pathogens such as Newcastle disease virus etc.The sensitivity of this assay was about 50pg of total RNA of AIV.Our data showed that this method was simple and feasible for rapid identification and subtyping of AIV subtype H_9 and H_5 at a single reaction.In addition,both diagnostic amplicons for AIV subtype H_9 and H_5 contained the cleavage site sequence of HA gene,from which the amino acid sequence might be deduced to predict the potential virulence of AIV.