Influenza viruses existing in birds continue to be a source of a diverse combination of antigenic subtypes, including 16 hemagglutinin (HA) and 9 neuraminidase (NA), and represent a vast reservoir of new antigens to which the human population is naive [1,2](1)) (Seasonal influenza epidemics constitute a major public health problem, accounting for five million serious cases worldwide [1 ](4))(Each year, influenza A types cause human epidemics responsible for significant mortality and morbidity, particularly in high-risk groups, such as infants, the elderly, and immunocompromised individuals. (5)).(3)) (Influenza virus is one of the most devastating viral diseases due to its high contagiousness and easy spread. aerosol and causes acute viral respiratory illness and mortality in susceptible groups. In order to prevent the spread of seasonal or pandemic influenza epidemics, vaccination is a powerful and cost-effective means [1](15)) (Protection against the influenza virus is mainly. mediated by antibodies against viral hemagglutinin (HA) [2,3]), HA is the main surface glycoprotein of the virion and is responsible for the attachment and penetration of viral particles into cells during initial stages of infection.((5)( (8)) (Effective prophylactic influenza vaccines produce effective HA-specific systemic antibodies, which can bind to the virus and inhibit early events of infection by the influenza virus (6)) Different types of influenza vaccines such as subunit [7-10], attenuated [11,12] and inactivated influenza vaccines [14] are available, although inactivated vaccines are the most widely used on a commercial scale [6]. .. middle of paper...... baculovirus stock P1 In Sf-900 Medium III was prepared, if necessary. To do this, 0.25 ml of baculovirus stock was diluted sequentially in 2.25 ml of Sf-900 medium. Dilutions 10-4 to 10-8 were used in our test. , immediately replace with 1 ml of the appropriate viral dilution and incubate for 1 hour at room temperature. Plate medium containing 12.5 ml of 4% low melting point agarose and 37.5 ml of Sf-900III was prepared and incubated in a water bath at 40°C until use. After one hour of incubation, the virus-containing medium from the wells is removed and replaced with 2 ml of plating medium. Allow the agarose to layer at room temperature until it hardens. The plates are incubated in a humidified incubator at 27 °C for 7 to 10 days. After viewing the plaques, the plaques were stained with 0.5 ml of neutral red solution (1 mg/ml). (21)