Effects Of Recombinant Avian Influenza Virus (H5N1 Subtype, Re-5 Strain) By Different Treatment And Different Times Of Preservation

This study uses production poisonous kinds of recombinant avian influenza virus (H5N1 subtype, Re-5 strain) strains of a certain percentage of diluted, inoculated in 11-day-old well-developed non-immune health of chick embryo cysts, each embryo 0.2ml, in the 10000 production in the region, the temperature 36℃, humidity 70% of the conditions for the continuation incubated for 72 hours, whichever is home of live chicken0~ 4℃refrigerator cool, 12 hours later, harvested chicken embryo allantoic fluid, as the untreated antigen. Untreated antigen through the centrifuge centrifuge, as the centrifugal antigen. After centrifugation, the antigen will be a filtration membrane pack twice the concentration, as the antigen concentration. Through the preparation of the untreated, centrifugal purified, concentrated sample of purified antigen were carried out to clarify the degree of pre-inactivated, the virus infected half of the amount of (EID50) and the price of chicken red blood cell agglutination (HA) of the detection tests to determine the antigen by centrifugation, concentrated Purification of the quality of treatment changes. Then, the inactivated agents were used formaldehyde solution 1.5‰, 2.0‰, 2.5‰inactivated, temperature 37℃inactivated 24 hours, by inactivated antigen inactivated sterile samples for testing, inactivated test, HA test inactivated detection tests to determine the untreated antigen, centrifugation purified antigens and inactivated concentrated purified antigens complete a minimum of formaldehyde concentration. Finally, each group with the lowest concentration of formaldehyde inactivated antigens of qualified packaging and sealing wrap is good, for 0~ 4℃preservation test, were saved by 1 month, 2 months, 3 months, 4 months, five months later, the HA test and make an oil-emulsion preparation of seedlings, seedlings of the oil emulsion appearance, properties, viscosity, stability, physical properties testing and 20-day-old non-immune chickens safety testing and 5-week-old non-immune HI antibody detection of immune chickens to determine the untreated antigen, centrifugal concentration of purified antigens and the quality of the purified antigen titer stability. Through the preparation of the untreated, centrifugal purification, concentration, purification of the antigen to clarify the degree of virus infection in half the amount of (EID50) and the price of chicken red blood cell agglutination (HA) test comparison of test results obtained: Antigen After centrifugation, concentration, purification , not only the appearance of significant improvement in quality and potency are greatly enhanced. The enrichment process to clarify the degree of antigen than untreated antigen, less than the centrifugal antigen; HA and centrifugal treatment compared with untreated antigen titer increased by one; EID50 than the centrifugal antigen EID50 increase of 0.2 over the unprocessed antigens increased 0.7. Through untreated, centrifugal purification, concentration, purification antigen 1.5‰, 2.0‰, 2.5‰formaldehyde solution inactivated and inactivated sterile after the inspection, inspection and HA test inactivated comparison of test results obtained: untreated antigen completely inactivated the lowest formaldehyde concentration of 2.5‰, the lowest centrifugal completely inactivated antigen concentration of formaldehyde solution, 1.5‰, the lowest concentration completely inactivated antigen concentration of formaldehyde solution 2.0‰, 2.5‰formaldehyde solution through untreated inactivated antigen, 1.5‰formaldehyde solution inactivated centrifugal antigens and 2.0‰enrichment of antigen inactivated formaldehyde solution to save one month, two months, three, four, five HA months of testing and preparation of the physical properties of oil-emulsion postemergence testing, safety testing and detection of immune antibody HI comparison of test results obtained: In the five months to save time, quality and stability of concentrated antigen, the immune good effect. HA antigen concentration than the average height of a centrifugal antigen titer, compared with an average of 3.6 untreated antigen titer; prepared Miao oil emulsion stability, security, body-immunized chickens produced HI antibody antigen than the average height of 1.08 centrifuge titer was higher than the average of 3.36 untreated antigen titer, and the immune antibody and tidy. The resulting: The centrifuge and enrichment technology can improve the quality of the avian influenza virus antigen, while antigen concentration doubling the size of antigen for the future there are plans to reserve, inactivated polyvalent Miao Miao and multi-joint production of laying a solid basis.