Preparation method of divalent inactivated vaccines for duck virus hepatitis

Abstract

The invention relates to a preparation method of divalent inactivated vaccines for duck virus hepatitis. According to the invention, the preparation method comprises the following steps of: screening duck virus hepatitis type I YBH1 strains and novel YBHX strain vaccines with good immunogenicity from preponderant popular strains so as to prepare strains, respectively inoculating the vaccines on a chicken embryo and a duck embryo, respectively harvesting embryoid bodies and embryoid fluid of dead embryos, collecting a viral solution after the embryoid bodies and the embryoid fluid are ground and frozen-thawed, and adding an oil adjuvant for mixing and emulsifying to obtain the vaccines after the viral solution is carried out ultrafiltration and concentration and is inactivated through a formaldehyde solution. The vaccines can carry out immunization for animals and can simultaneously prevent the duck virus hepatitis caused by type I duck virus hepatitis and novel duck virus hepatitis, and the divalent inactivated vaccines for the duck virus hepatitis have the advantages of high efficiency, good safety and high protective ratio and the like.

Description

technical field

[0001] The invention relates to a preparation method of a duck viral hepatitis bivalent inactivated vaccine, belonging to the field of veterinary biological products. Background technique

[0002] Duck viral hepatitis (DVH) is an acute, highly fatal, contagious disease of ducklings caused by duck hepatitis virus (DHV) with liver as the main pathological change. Clinical manifestations include neurological symptoms such as spasms, convulsions, and opisthotonus, and liver swelling and hemorrhage are characteristic lesions. Ducklings aged 1 to 3 weeks are susceptible to severe disease and spread rapidly. Ducklings usually die within 1 hour after symptoms appear, and the mortality rate of 10-day-old ducklings after infection is 90% to 95%. It has become a serious hazard to duck breeding. one of the major diseases of the industry.

[0003] Duck hepatitis virus (DHV) serotype DHV has 4 independent serotypes, type 1, type 2, type 3 and new type. Fan Shucai et al....

Examples

Embodiment 1

[0047] (1) Vaccine (immunogen) preparation:

[0048] 1 Preparation of virus solution for seedling production

[0049] Preparation of YBH1 strain virus liquid: Dilute the virus strain YBH1 used for production to 100 times, inoculate 100 10-day-old SPF chicken embryos into the allantoic cavity, 0.2ml per embryo, incubate at 36-37°C, and inspect the embryos twice a day. Chicken embryos that died within 48 to 120 hours after inoculation were placed at 2 to 8°C for 4 to 12 hours, and allantoic fluid, amniotic fluid and embryo bodies were collected, and the heads and limbs were removed from the embryo bodies. Homogenize with embryo fluid, freeze and thaw three times, centrifuge at 4000r / min for 30min, take the supernatant and mix it in a sterile container, store at 2-8°C. (See Table 1).

[0050] Preparation of YBHX strain virus liquid Dilute the virus species YBHX strain used for production by 100 times, inoculate 100 susceptible duck embryos of 12-13 days old in the allantoic cav...

Embodiment 2

[0071] ——Inspection of finished vaccines

[0072] 1 character

[0073] (1) Appearance Milky white emulsion.

[0074] (2) Dosage form Water-in-oil type, take a clean straw, absorb a small amount of vaccine and drop it in cold water, and it will not spread in the form of oil droplets.

[0075] (3) Stability Take 10ml of the vaccine and put it into a centrifuge tube, centrifuge at 3000r / min for 15 minutes, no water phase will precipitate out at the bottom of the tube.

[0076] (4) Viscosity is 61.6 cp according to the appendix of the current version of "Chinese Veterinary Pharmacopoeia".

[0077] 2 Inspection of the filling volume is carried out according to the appendix of the current version of "Chinese Veterinary Pharmacopoeia", and it meets the standard.

[0078] 3 Sterility test According to the appendix of the current version of "Chinese Veterinary Pharmacopoeia", the sterile growth is carried out.

[0079] 4 For safety inspection, 10 SPF chickens aged 21 to 35 days wer...

technical field

[0001] The invention relates to a preparation method of a duck viral hepatitis bivalent inactivated vaccine, belonging to the field of veterinary biological products. Background technique

[0002] Duck viral hepatitis (DVH) is an acute, highly fatal, contagious disease of ducklings caused by duck hepatitis virus (DHV) with liver as the main pathological change. Clinical manifestations include neurological symptoms such as spasms, convulsions, and opisthotonus, and liver swelling and hemorrhage are characteristic lesions. Ducklings aged 1 to 3 weeks are susceptible to severe disease and spread rapidly. Ducklings usually die within 1 hour after symptoms appear, and the mortality rate of 10-day-old ducklings after infection is 90% to 95%. It has become a serious hazard to duck breeding. one of the major diseases of the industry.

[0003] Duck hepatitis virus (DHV) serotype DHV has 4 independent serotypes, type 1, type 2, type 3 and new type. Fan Shucai et al....

Examples

Embodiment 1

[0047] (1) Vaccine (immunogen) preparation:

[0048] 1 Preparation of virus solution for seedling production

[0049] Preparation of YBH1 strain virus liquid: Dilute the virus strain YBH1 used for production to 100 times, inoculate 100 10-day-old SPF chicken embryos into the allantoic cavity, 0.2ml per embryo, incubate at 36-37°C, and inspect the embryos twice a day. Chicken embryos that died within 48 to 120 hours after inoculation were placed at 2 to 8°C for 4 to 12 hours, and allantoic fluid, amniotic fluid and embryo bodies were collected, and the heads and limbs were removed from the embryo bodies. Homogenize with embryo fluid, freeze and thaw three times, centrifuge at 4000r / min for 30min, take the supernatant and mix it in a sterile container, store at 2-8°C. (See Table 1).

[0050] Preparation of YBHX strain virus liquid Dilute the virus species YBHX strain used for production by 100 times, inoculate 100 susceptible duck embryos of 12-13 days old in the allantoic cav...

Embodiment 2

[0071] ——Inspection of finished vaccines

[0072] 1 character

[0073] (1) Appearance Milky white emulsion.

[0074] (2) Dosage form Water-in-oil type, take a clean straw, absorb a small amount of vaccine and drop it in cold water, and it will not spread in the form of oil droplets.

[0075] (3) Stability Take 10ml of the vaccine and put it into a centrifuge tube, centrifuge at 3000r / min for 15 minutes, no water phase will precipitate out at the bottom of the tube.

[0076] (4) Viscosity is 61.6 cp according to the appendix of the current version of "Chinese Veterinary Pharmacopoeia".

[0077] 2 Inspection of the filling volume is carried out according to the appendix of the current version of "Chinese Veterinary Pharmacopoeia", and it meets the standard.

[0078] 3 Sterility test According to the appendix of the current version of "Chinese Veterinary Pharmacopoeia", the sterile growth is carried out.

[0079] 4 For safety inspection, 10 SPF chickens aged 21 to 35 days wer...

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