Combined inactivate vaccine for egg drop syndrome and avian encephalomlris and preparation method of combined inactivate vaccine

A dual inactivated vaccine and egg reduction syndrome technology, applied in the field of bioengineering, can solve problems such as losses in the chicken industry, and achieve the effects of good immunogenicity, high antibody level, and reduced dosage

Pending Publication Date: 2016-12-07
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the report of this disease occurred in our country in the mid-1980s, it has occurred

Method used

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  • Combined inactivate vaccine for egg drop syndrome and avian encephalomlris and preparation method of combined inactivate vaccine
  • Combined inactivate vaccine for egg drop syndrome and avian encephalomlris and preparation method of combined inactivate vaccine
  • Combined inactivate vaccine for egg drop syndrome and avian encephalomlris and preparation method of combined inactivate vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Screening and Identification of Avian Encephalomyelitis HM08 Strain

[0027] 1. Virus Isolation

[0028] Kill the sick chicks with the pathological characteristics of avian encephalomyelitis, aseptically collect the brain tissues of the sick and dead chicks, mix them, freeze and thaw them three times, grind them in a sterile container, and make a 1:3 suspension with PBS buffer , add penicillin and streptomycin at 1000IU / ml, centrifuge at 3000r / min for 20 minutes, take the supernatant, inoculate 10 6-day-old SPF chicken embryos through the yolk sac, 0.2ml per embryo, seal and place at 37.0 Incubate at ℃. After hatching to 18 days old, observe the lesions of the chicken embryos. If no obvious lesions appear within 12 days after inoculation, take out the embryonic brain, homogenate, make a suspension with the same batch of allantoic fluid or PBS buffer at a ratio of 1:3, and put it out. Freeze and thaw three times in the container of bacteria, centrifuge at 300...

Embodiment 2

[0065] Example 2 Preparation of Egg Drop Syndrome and Avian Encephalomyelitis Dual Inactivated Vaccine

[0066] 1. Source of poisonous seeds for seedling production:

[0067] Chicken Egg Drop Syndrome Virus AV127 strain was purchased from China Veterinary Drug Control Institute.

[0068] Avian encephalomyelitis virus HM08 strain, preservation number: CGMCC NO.7759.

[0069] 2. Preparation of virus liquid:

[0070] (1) Preparation of egg drop syndrome virus AV127 strain virus fluid: inoculate susceptible duck embryos with chicken egg drop syndrome virus AV127 strain, harvest duck embryo fluid as chicken egg drop syndrome virus AV127 strain virus fluid.

[0071] The specific method is as follows:

[0072] Inoculation: take chicken egg drop syndrome virus AV127 strain, make appropriate dilution with sterilized physiological saline (such as 10 -1 or 10 -2 ), inoculate 8-10-day-old susceptible duck embryos in the allantoic cavity, 0.1ml per embryo, seal the pinhole after inocu...

Embodiment 3

[0105] Example 3 Double-dose Inoculation Safety Test of Chicken Egg Drop Syndrome and Avian Encephalomyelitis Dual Inactivated Vaccine

[0106] Prepare 3 batches of chicken egg drop syndrome and avian encephalomyelitis dual inactivated vaccines according to Example 2 (batch numbers: 20130328, 20130427, 20130909, the content of AV127 strains in the 0.3ml dual inactivated vaccines is 2000HAU, and the content of HM08 strains is 10 5.0 EID 50 ).

[0107] The research objects were 7-day-old SPF chickens (purchased from Beijing Meria Weitong Experimental Animal Co., Ltd.) and 7-day-old Sanhuang broiler chickens (purchased from Nanjing Shifosi Chicken Farm). After each batch of chickens was bought back, they were adapted for a few days to check the general condition and clinical disease status of the chickens. Weak chickens were eliminated and healthy chickens were selected to enter the test. SPF chickens were double-dose (0.6ml / only) subcutaneously injected three batches of chick...

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Abstract

The invention provides a combined inactivate vaccine for the egg drop syndrome and avian encephalomlris and a preparation method of the combined inactivate vaccine, and relates to the field of bioengineering. The combined inactivate vaccine contains an inactivate egg drop syndrome virus AV127 and an avian encephalomlris virus HM08 strain, the conservation number of the HM08 strain is CGMCC No.7759. The preparation method of the vaccine is as follows: two virus strains are respectively inoculated on a susceptible duck embryo and a chick embryo SPF to obtain an egg drop syndrome virus solution and an avian encephalomlris virus solution; the inactivate virus solutions are mixed and then evenly mixed with a Twain-80 to prepare an aqueous phase, the aqueous phase and an oil phase are evenly mixed to obtain the combined inactivate vaccine. The HM08 strain propagates in the chick embryo SPF, the content of 0.2ml of the virus solution is more than or equal to 107.0EID50, the immunogenicity is good, the produced antibody level is high, the cross protection is good, and the combined inactivate vaccine produces a high-level antibody for a long persistent period.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a dual inactivated vaccine for egg drop syndrome and avian encephalomyelitis and a preparation method thereof. Background technique [0002] Egg drop syndrome (Egg Drop Syndrome, EDS) is caused by adenovirus. Decline, a large number of soft-shell eggs and deformed eggs appear, and the eggshell color becomes lighter, which brings great economic losses to production. Since the Dutch scientist van Eck first reported the disease in 1976, many countries and regions have reported the prevalence of the disease. Since egg drop syndrome was discovered in broiler breeder flocks in my country in the late 1980s, the disease has been prevalent in many areas. [0003] Avian encephalomyelitis (Avian Encephalomyelitis, AE), also known as epidemic tremor, is caused by avian encephalomyelitis virus (AEV), which mainly harms chicks under 4 weeks old, in order to invade the central nervous system of ...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/235A61K39/125A61P31/14
CPCA61K39/12A61K2039/521A61K2039/70C12N2710/10034C12N2770/32334
Inventor 朱亚露揭鸿英于漾何家惠侯继波毕志香赵阳唐应华
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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