CN102526727B - Anti-bird-flue egg yolk antibody injection and application thereof - Google Patents
Anti-bird-flue egg yolk antibody injection and application thereof Download PDFInfo
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- CN102526727B CN102526727B CN201210000909.8A CN201210000909A CN102526727B CN 102526727 B CN102526727 B CN 102526727B CN 201210000909 A CN201210000909 A CN 201210000909A CN 102526727 B CN102526727 B CN 102526727B
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- 210000002969 egg yolk Anatomy 0.000 title claims abstract description 91
- 238000002347 injection Methods 0.000 title claims abstract description 84
- 239000007924 injection Substances 0.000 title claims abstract description 84
- 102000002322 Egg Proteins Human genes 0.000 title claims abstract description 59
- 108010000912 Egg Proteins Proteins 0.000 title claims abstract description 59
- 235000013345 egg yolk Nutrition 0.000 title claims abstract description 54
- 238000002360 preparation method Methods 0.000 claims abstract description 24
- 206010064097 avian influenza Diseases 0.000 claims description 89
- 208000002979 Influenza in Birds Diseases 0.000 claims description 61
- 230000036039 immunity Effects 0.000 claims description 37
- 206010015856 Extrasystoles Diseases 0.000 claims description 21
- 230000003053 immunization Effects 0.000 claims description 20
- 238000002649 immunization Methods 0.000 claims description 20
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- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 14
- 229960003971 influenza vaccine Drugs 0.000 claims description 14
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- 230000003068 static effect Effects 0.000 claims description 10
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- 239000003795 chemical substances by application Substances 0.000 claims description 7
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 6
- 230000003260 anti-sepsis Effects 0.000 claims description 6
- 238000011081 inoculation Methods 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 5
- 210000003278 egg shell Anatomy 0.000 claims description 5
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- 238000000926 separation method Methods 0.000 claims description 5
- 239000001632 sodium acetate Substances 0.000 claims description 5
- 235000017281 sodium acetate Nutrition 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000008215 water for injection Substances 0.000 claims description 5
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 claims description 3
- 229940033663 thimerosal Drugs 0.000 claims description 3
- 241000287828 Gallus gallus Species 0.000 abstract description 36
- 238000010255 intramuscular injection Methods 0.000 abstract description 8
- 239000007927 intramuscular injection Substances 0.000 abstract description 8
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- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses an anti-bird-flue egg yolk antibody injection. A preparation method of the injection comprises the following steps of: preparing an anti-bird-flue antibody; preparing a bird flue egg yolk antibody injection, and the like. The injection can be applied to prevention of broiler bird flue, and is applied to intramuscular injection in the amount of 0.2-0.5mL for every 1kg of broiler. The injection is used for preventing bird flue by raising the bird flue antibody levels in broilers, has the characteristics of nontoxicity harmlessness, no residue and environmental friendliness when used on broilers, and can be used for improving the poultry quality. The anti-bird-flue egg yolk antibody injection disclosed by the invention is further applied to prevention of a broiler bird flue disease.
Description
Technical field
The invention belongs to immunological technique field, relate to a kind of yolk antibody preparation and application thereof, specifically a kind of Anti-bird-flue egg yolk antibody injection and uses thereof.
Background technology
Bird flu is that the one of the poultry that causes of influenza virus is acute, height lethal infectious disease, and primary disease is mainly to prevent by immunity inoculation avian influenza vaccine at present.But because broiler chicken breeding cycle is shorter, avian influenza vaccine is inactivated vaccine, before delivering for sale, not yet absorb completely, in muscle, form many lesser tubercle, will inevitably affect the quality of Carnis Gallus domesticus.
Summary of the invention
The technical problem to be solved in the present invention, is to provide a kind of Anti-bird-flue egg yolk antibody injection and uses thereof.
The prepared anti-avian influenza yolk antibody of the present invention is immunoglobulin, belongs to protein, and molecular weight is easily absorbed by body after making injection enter in body, and muscle is difficult for producing tuberosity; By the antibody horizontal that improves broiler chicken body bird flu, carry out birds flu-preventing, Anti-bird-flue egg yolk antibody injection is used on broiler chicken, and the efficacy of a drug is more direct, drug effect is faster, birds flu-preventing, easy to operate, flexible, effective, and contribute to the raising of chicken meat product quality.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
1, agents useful for same
Ammonium sulfate (analytical pure, purity >=99.0%); Thimerosal (analytical pure, purity >=98.0%); New stream H9 bigeminy oil seepage (manufacturer: Yangzhou Brunswick biological engineering company limited); H 5 N 1 avian influenza-4-5 oil seepage (manufacturer: Harbin Wei Ke biotechnology development company); Bird flu H9(manufacturer: Yangzhou Brunswick biological engineering company limited)
2, preparation method
A kind of Anti-bird-flue egg yolk antibody injection, the preparation method of this injection is carried out according to following steps order:
1. induction produces anti-avian influenza antibody
By immune programme for children immunity bird inlay, carry out inoculation with avian influenza vaccine, induction of immunity hen produces anti-avian influenza antibody;
2. the preparation of avian influenza egg yolk antibodies injection
Collect after immune bird inlay yolk, then yolk antibody is extracted and purification, obtain Anti-bird-flue egg yolk antibody injection.
As a kind of restriction of the present invention, described immune programme for children is:
When I, maternal antibody are low, the immunization method of different days section:
7 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL; 15 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
When II, maternal antibody are high, the immunization method of different days section:
14 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL;
21 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
40 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 0.5mL;
50 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.5mL;
110 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 1mL;
130 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 1mL;
III, every 60 days, use H 5 N 1 avian influenza-4-5 oil seepage to carry out booster immunization one time later, every 80 days, use bird flu H9 to carry out booster immunization one time, bird inlay is carried out to immunity, then extract immune bird inlay yolk preparation.
As another kind of restriction of the present invention, the preparation method of described extraction purification and injection is carried out according to following steps order:
(1) separation of yolk
Height is exempted to egg sterilization, under aseptic condition, break eggshell, isolate yolk;
(2) extraction of supernatant
Egg yolk is poured in sterilized container, and adding buffer to regulate pH value is 5.5-7.5, and after homogenate, 0-4 ℃ of static 3-4h or spend the night, collects supernatant;
(3) collection of precipitation
In supernatant, add the ammonium sulfate of 25-40%, the centrifugal 30min of 5000r/min, 0-4 ℃ of static 3-4h or spend the night, collecting precipitation;
(4) preparation of injection
10% the water for injection that adds former supernatant volume, makes resolution of precipitate, adds 0.01% antisepsis antistaling agent, and 0.22 micron of membrane filtration degerming, makes injection, freezing preservation.
As further restriction, described buffer is acetic acid and sodium acetate.
The present invention also has a kind of restriction, and described antisepsis antistaling agent is thimerosal.
The present invention also provides a kind of purposes of above-mentioned Anti-bird-flue egg yolk antibody injection, is about to described Anti-bird-flue egg yolk antibody injection for preventing broiler chicken bird flu.
Described Anti-bird-flue egg yolk antibody injection carries out intramuscular injection according to the amount of the every 1kg body weight of broiler chicken 0.2-0.5mL.
The application of Anti-bird-flue egg yolk antibody injection: for prepared above-mentioned Anti-bird-flue egg yolk antibody injection, detect effect, by contrast test and employing hemagglutination inhibition test, measure antibody titer, effect to prepared injection is evaluated, and wherein detection method is with reference to GB: the hemagglutination inhibition test of GB/T 14926.54-2001 experimental animal.
Contrast test: 120 broiler chicken are divided into three groups, be respectively: 25 ages in days are not injected this Anti-bird-flue egg yolk antibody injection and immune avian influenza vaccine, 25 ages in days are not injected this Anti-bird-flue egg yolk antibody injection and three groups of the broiler chicken of immune avian influenza vaccine and 25 age in days broiler chicken are not at 30 age in days counteracting toxic substances bird flu antigen 0.2mL/ only, other raising condition is the same, observe to delivering for sale, result is as shown in the table:
Wherein: intramuscular injection "-" represents not inject any injection and vaccine; Antibody titer "-" indicates without antibody titer.
Comparative test result represents: broiler chicken is injected the chicken group of this Anti-bird-flue egg yolk antibody injection and immune avian influenza vaccine, and chicken group situation is without significant difference, but can improve meat quality.This Anti-bird-flue egg yolk antibody injection provided by the present invention has certain preventive effect and without any side effects to broiler chicken bird flu disease.
The mensuration of wherein tiring is by Anti-bird-flue egg yolk antibody injection, detects the yolk antibody of injection tire by hemagglutination inhibition test (operational approach is carried out according to GB/T18936-2003), and all >=8log2 that tires of finished product, is qualified products after testing.
Owing to having adopted above-mentioned technical scheme, the present invention compared with prior art, obtained technological progress is: the preparation technology of Anti-bird-flue egg yolk antibody injection is simple, cost is low, nontoxic, harmless, noresidue, pollution-free, and easy to use, be all that the antibody horizontal by improving broiler chicken body bird flu carrys out birds flu-preventing disease; The prepared anti-avian influenza yolk antibody of the present invention is immunoglobulin, belongs to protein, and molecular weight is easily absorbed by body after making injection enter in body, and muscle is difficult for producing tuberosity; Anti-bird-flue egg yolk antibody injection provided by the present invention is used on broiler chicken, and the efficacy of a drug is more directly, drug effect is faster, compared with prior art has birds flu-preventing, easy to operate, flexible, effective feature, and contribute to the raising of chicken meat product quality.
Method of the present invention, for the preparation of Anti-bird-flue egg yolk antibody injection, is applied to the prevention of broiler chicken bird flu disease further.
The present invention is described in further detail below in conjunction with specific embodiment.
The specific embodiment
embodiment 1
A kind of Anti-bird-flue egg yolk antibody injection, the preparation method of this injection is carried out according to following steps order:
1. induction produces anti-avian influenza antibody
By immune programme for children immunity bird inlay, carry out inoculation with avian influenza vaccine, induction of immunity hen produces anti-avian influenza antibody.Wherein, immune programme for children is:
When I, maternal antibody are low, the immunization method of different days section:
7 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL; 15 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
When II, maternal antibody are high, the immunization method of different days section:
14 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL;
21 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
40 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 0.5mL;
50 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.5mL;
110 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 1mL;
130 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 1mL;
III, every 60 days, use H 5 N 1 avian influenza-4-5 oil seepage to carry out booster immunization one time later, every 80 days, use bird flu H9 to carry out booster immunization one time, bird inlay is carried out to immunity, then in subsequent step, extract immune bird inlay yolk Dispersal risk.
2. the preparation of avian influenza egg yolk antibodies injection
Collect after immune bird inlay yolk, then yolk antibody is extracted and purification, then prepare injection through conventional method, make a kind of Anti-bird-flue egg yolk antibody injection.
Wherein, " extracting and purification " is to carry out according to following steps order:
(1) separation of yolk
Height is exempted to egg sterilization, under aseptic condition, break eggshell, isolate yolk;
(2) extraction of supernatant
Egg yolk is poured in sterilized container, added the buffer of acetic acid and sodium acetate, regulating pH value is 5.5; After homogenate, 4 ℃ of static 3h or spend the night, collect supernatant;
(3) collection of precipitation
In supernatant, add 25% ammonium sulfate, the centrifugal 30min of 5000r/min, 0 ℃ of static 3h, collecting precipitation.
The preparation of injection: add 10% water for injection of supernatant volume, make resolution of precipitate, add 0.01% antisepsis antistaling agent, 0.22 micron of membrane filtration degerming, makes injection, freezing preservation.
The present embodiment, also by prepared above-mentioned Anti-bird-flue egg yolk antibody injection, is applied to prevention broiler chicken bird flu, and concrete mode is to carry out intramuscular injection according to the amount of the every 1kg body weight of broiler chicken 0.5mL.
embodiment 2
A kind of Anti-bird-flue egg yolk antibody injection, the preparation method of this injection is carried out according to following steps order:
1. induction produces anti-avian influenza antibody
By immune programme for children immunity bird inlay, carry out inoculation with avian influenza vaccine, induction of immunity hen produces anti-avian influenza antibody.Wherein, immune programme for children is:
When I, maternal antibody are low, the immunization method of different days section:
7 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL; 15 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
When II, maternal antibody are high, the immunization method of different days section:
14 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL;
21 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
40 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 0.5mL;
50 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.5mL;
110 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 1mL;
130 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 1mL;
III, every 60 days, use H 5 N 1 avian influenza-4-5 oil seepage to carry out booster immunization one time later, every 80 days, use bird flu H9 to carry out booster immunization one time, bird inlay is carried out to immunity.
2. the preparation of avian influenza egg yolk antibodies injection
Collect after immune bird inlay yolk, then yolk antibody is extracted and purification, then prepare injection through conventional method, make a kind of Anti-bird-flue egg yolk antibody injection.
Wherein, in yolk antibody injection, the method for the extraction purification of yolk antibody is carried out according to following steps order:
(1) separation of yolk
Height is exempted to egg sterilization, under aseptic condition, break eggshell, isolate yolk;
(2) extraction of supernatant
Egg yolk is poured in sterilized container, added the buffer of acetic acid and sodium acetate, regulate pH value to be 7.5,0 ℃ of static 4h or to spend the night, collect supernatant;
(3) collection of precipitation
In supernatant, add 40% ammonium sulfate, the centrifugal 30min of 5000r/min, 4 ℃ of static 4h or spend the night, collecting precipitation.
The preparation of injection: add 10% water for injection of former supernatant volume, make resolution of precipitate, add 0.01% antisepsis antistaling agent, 0.22 micron of membrane filtration degerming, makes injection, freezing preservation.
The present embodiment also provides a kind of purposes of above-mentioned Anti-bird-flue egg yolk antibody injection, uses it for prevention broiler chicken bird flu, according to the amount of the every 1kg body weight of broiler chicken 0.3mL, carries out intramuscular injection.
embodiment 3
A kind of Anti-bird-flue egg yolk antibody injection, the preparation method of this injection is carried out according to following steps order:
1. induction produces anti-avian influenza antibody
By immune programme for children immunity bird inlay, carry out inoculation with avian influenza vaccine, induction of immunity hen produces anti-avian influenza antibody.Wherein, immune programme for children is:
When I, maternal antibody are low, the immunization method of different days section:
7 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL; 15 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
When II, maternal antibody are high, the immunization method of different days section:
14 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL;
21 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
40 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 0.5mL;
50 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.5mL;
110 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 1mL;
130 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 1mL;
III, every 60 days, use H 5 N 1 avian influenza-4-5 oil seepage to carry out booster immunization one time later, every 80 days, use bird flu H9 to carry out booster immunization one time, bird inlay is carried out to immunity.
2. the preparation of Anti-bird-flue egg yolk antibody injection
Collect after immune bird inlay yolk, then yolk antibody is extracted and purification, then prepare injection through conventional method, make a kind of Anti-bird-flue egg yolk antibody injection.
Wherein, in yolk antibody injection, the extraction purification of yolk antibody is to carry out according to following steps order:
(1) separation of yolk
Height is exempted to egg sterilization, under aseptic condition, break eggshell, isolate yolk;
(2) extraction of supernatant
Egg yolk is poured in sterilized container, added the buffer of acetic acid and sodium acetate, regulate pH value to be 6.5,2 ℃ of static 4h or to spend the night, collect supernatant;
(3) collection of precipitation
In supernatant, add 37% ammonium sulfate, the centrifugal 30min of 5000r/min, 3 ℃ of static 3h or spend the night, collecting precipitation.
The preparation of injection: add 10% water for injection of former supernatant volume, make resolution of precipitate, add 0.01% antisepsis antistaling agent, 0.22 micron of membrane filtration degerming, makes injection, freezing preservation.
The present embodiment is also applied to above-mentioned Anti-bird-flue egg yolk antibody injection prevention broiler chicken bird flu, according to the amount of the every 1kg body weight of broiler chicken 0.2mL, carries out intramuscular injection.
embodiment 4the application of the prepared a kind of Anti-bird-flue egg yolk antibody injection of embodiment 1-3
The application of Anti-bird-flue egg yolk antibody injection: for the prepared above-mentioned Anti-bird-flue egg yolk antibody injection of embodiment 1-3, detect effect, by comparative, test and adopt hemagglutination inhibition test to measure antibody titer, effect to prepared injection is evaluated, and wherein detection method is with reference to GB: the hemagglutination inhibition test of GB/T 14926.54-2001 experimental animal.
Contrast test: 120 broiler chicken are divided into three groups, be respectively: 25 ages in days do not inject this Anti-bird-flue egg yolk antibody injection and immune avian influenza vaccine, 25 ages in days are not injected this Anti-bird-flue egg yolk antibody injection and not three groups of the broiler chicken of immune avian influenza vaccine and 25 age in days broiler chicken 0.2mL/ chicken of 30 age in days counteracting toxic substances bird flu antigens, other raising condition is identical with the use of counteracting toxic substances antigen, observe to delivering for sale, result is as shown in table 1:
Wherein: intramuscular injection "-" represents not inject any injection and vaccine; Antibody titer "-" indicates without antibody titer.
[0037]comparative test result represents: broiler chicken is injected the chicken group of this Anti-bird-flue egg yolk antibody injection and immune avian influenza vaccine, and chicken group situation is without big difference, but can improve meat quality.This Anti-bird-flue egg yolk antibody injection providing has certain preventive effect and without any side effects to broiler chicken bird flu disease.
The mensuration of wherein tiring is by Anti-bird-flue egg yolk antibody injection, detects the yolk antibody of injection tire by hemagglutination inhibition test (operational approach GB/T18936-2003 execution), and all >=8log2 that tires of finished product, is qualified products after testing.
embodiment 5the quality standard of the prepared anti-avian influenza yolk antibody of embodiment 1-3:
[character] this product is light yellow to yellow liquid, has been long placed in precipitation.
[inspection] pH value should be 5.0～7.0.
Asepticly by < < Chinese veterinary pharmacopoeia > >, undertaken, should be without bacterial growth.
Detection of mycoplasma is undertaken by < < Chinese veterinary pharmacopoeia > >, should grow without mycoplasma.
Exogenous virus detects and is undertaken by < < Chinese veterinary pharmacopoeia > >, should conform with the regulations.
[titration] is by Anti-bird-flue egg yolk antibody injection by the mensuration of wherein tiring, and detects the yolk antibody of injection tire by hemagglutination inhibition test (operational approach GB/T18936-2003 execution), and the tire >=8log2 of finished product is qualified products.
[effect and purposes] is for the prevention of broiler chicken avian influenza.
The every 1kg body weight of [usage and consumption] intramuscular injection 0.2-0.5mL, once a day, can be used in conjunction 2-3 days.
[points for attention] 1, this product this product are passive immunity, no longer inject avian influenza vaccine in 7 days.
2, this product is oral invalid.
3, can mix use with antibiotic for animals.
4, the passive immunity protection period after this product injection is 5～7.
[storage] 2-8 ℃ preserved or room temperature preservation.
embodiment 6anti-bird-flue egg yolk antibody injection stability test
The concrete operations of anti-avian influenza yolk antibody stability test are as follows:
Accelerated test is placed 6 months under the condition of 25 ± 2 ℃ of temperature, relative humidity 60 ± 5%, in duration of test sampling in each month once, quality standard > > according to < < anti-avian influenza yolk antibody detects, and result is as shown in table 2:
Wherein, aseptic, detection of mycoplasma and exogenous virus detect test operation and all according to the examination criteria of the quality standard > > of < < anti-avian influenza yolk antibody, detect, and testing result all conforms with the regulations.
Claims (1)
1. an Anti-bird-flue egg yolk antibody injection,
it is characterized in that:the preparation method of this injection is carried out according to following steps order:
1. induction produces anti-avian influenza antibody
By immune programme for children, bird inlay is carried out to inoculation with avian influenza vaccine, induction of immunity hen produces anti-avian influenza antibody;
2. the preparation of avian influenza egg yolk antibodies injection
Collect after immune bird inlay yolk, then to yolk antibody extract and purification after, be prepared into Anti-bird-flue egg yolk antibody injection; Wherein:
A, described immune programme for children are:
When I, maternal antibody are low, the immunization method of different days section:
7 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL; 15 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
When II, maternal antibody are high, the immunization method of different days section:
14 ages in days: the new stream of immunity H9 bigeminy oil seepage 0.2mL;
21 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.3mL;
40 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 0.5mL;
50 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 0.5mL;
110 ages in days: immunity new stream H9 bigeminy oil seepage or bird flu H9 1mL;
130 ages in days: immune H 5 N 1 avian influenza-4-5 oil seepage 1mL;
III, every 60 days, use H 5 N 1 avian influenza-4-5 oil seepage to carry out booster immunization one time later, every 80 days, use bird flu H9 to carry out booster immunization one time, bird inlay is carried out to immunity, then extract immune bird inlay yolk preparation;
b,the preparation method of described extraction, purification and injection is carried out according to following steps order:
(1) separation of yolk
Height is exempted to egg sterilization, under aseptic condition, break eggshell, isolate yolk;
(2) extraction of supernatant
Egg yolk is poured in sterilized container, and adding buffer acetic acid and sodium acetate to regulate pH value is 5.5-7.5, and after homogenate, 0-4 ℃ of static 3-4h or spend the night, extracts supernatant;
(3) collection of precipitation
In supernatant, add the ammonium sulfate of 25-40%, the centrifugal 30min of 5000r/min, 0-4 ℃ of static 3-4h or spend the night, collecting precipitation;
(4) preparation of injection
10% the water for injection that adds supernatant volume, makes resolution of precipitate, adds 0.01% antisepsis antistaling agent thimerosal, and 0.22 micron of membrane filtration degerming, makes injection, freezing preservation.
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| CN1384119A (en) * | 2002-05-17 | 2002-12-11 | 重庆和润实业(集团)有限公司 | Composite yolk antibody for resisting fowl's viral blight and its prepn and application |
| CN1958608A (en) * | 2005-10-31 | 2007-05-09 | 雅臣药业集团（远东）有限公司 | Method for preparing specificity IgY of anti influenza of birds, and preparation |
| CN101732716A (en) * | 2008-11-04 | 2010-06-16 | 中国科学院动物研究所 | Antigen-antibody complex for preventing and/or treating avian influenza |
| CN102516389A (en) * | 2011-12-28 | 2012-06-27 | 黑龙江省农业科学院畜牧研究所 | Method for extracting avian influenza egg yolk antibodies |
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| CN1384119A (en) * | 2002-05-17 | 2002-12-11 | 重庆和润实业(集团)有限公司 | Composite yolk antibody for resisting fowl's viral blight and its prepn and application |
| CN1958608A (en) * | 2005-10-31 | 2007-05-09 | 雅臣药业集团（远东）有限公司 | Method for preparing specificity IgY of anti influenza of birds, and preparation |
| CN101732716A (en) * | 2008-11-04 | 2010-06-16 | 中国科学院动物研究所 | Antigen-antibody complex for preventing and/or treating avian influenza |
| CN102516389A (en) * | 2011-12-28 | 2012-06-27 | 黑龙江省农业科学院畜牧研究所 | Method for extracting avian influenza egg yolk antibodies |
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