CN103834619A - Duck virus hepatitis virus, duck virus hepatitis inactivated vaccine and preparation method of duck virus hepatitis inactivated vaccine - Google Patents
Duck virus hepatitis virus, duck virus hepatitis inactivated vaccine and preparation method of duck virus hepatitis inactivated vaccine Download PDFInfo
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- CN103834619A CN103834619A CN201310354525.0A CN201310354525A CN103834619A CN 103834619 A CN103834619 A CN 103834619A CN 201310354525 A CN201310354525 A CN 201310354525A CN 103834619 A CN103834619 A CN 103834619A
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Abstract
The invention provides a duck virus hepatitis virus, a duck virus hepatitis inactivated vaccine and a preparation method of the duck virus hepatitis inactivated vaccine. The duck virus hepatitis virus is preserved in China General Microbiological Culture Collection Center with the preservation number of CGMCC No.7804. The duck virus hepatitis virus has strong toxicity, good immunogenicity, and high toxicity evaluation, and the vaccine prepared by the duck virus hepatitis virus has good safety and long immune protection period.
Description
Technical field
The present invention relates to biological technical field, be specifically related to duck viral hepatitis virus, duck virus hepatitis inactivated vaccine and preparation method thereof.
Background technology
Duck viral hepatitis disease is acute, septic, the contagious disease of a kind of main infringement duckling.Duckling in these disease main infection 7 ages in days, is acute process after morbidity, and sickness rate can reach 100%, and mortality ratio can be up to 80～100%.Ducks more than 10 ages in days is the stealthy process that infects, and mortality ratio reduces along with the increase of age in days.Duck viral hepatitis disease has caused tremendous economic loss to supporting duck industry, becomes foster duck industry and endangers maximum epidemic disease.
This disease all has outburst all over the world, and often causes big area popular, is mainly to endanger 20 ages in days with interior duckling, and causes 7 ages in days with interior duck 100% Mortality.But for duckling, its immunity system is grown not perfect, there is many difficulties factor and the undesirable practical problems of immunne response in vaccine immunity in practice.Therefore, the yellow antibody prevention of preparation specificity high-immunity egg and control duck viral hepatitis disease, become the most economical effective means of controlling this sick outbreak of epidemic.
But, at present the domestic duck viral hepatitis having gone on the market high exempt from antibody exist unstable, the duck embryo of tiring can some homology disease of vertical transmission, there are the problems such as hidden danger in off quality, the yellow antibody producing technique of duck egg of duck embryo.The research of therefore, exempting from antibody about duck viral hepatitis is high need further improvement.
Summary of the invention
The present invention is intended to one of solve the problems of the technologies described above at least to a certain extent.For this reason, one object of the present invention is to propose a kind of duck viral hepatitis virus, duck virus hepatitis inactivated vaccine and preparation method thereof.
The following discovery of the present invention based on contriver completes: contriver is first from Shandong Mou Ya factory, gather ill duck liver organization, being equipped with sterilizing PBS grinds, filter the centrifugal supernatant liquor of getting afterwards, after the degerming of 0.22um filtering with microporous membrane, inoculation 11～13 age in days SPF chick embryo allantoic cavities, under 37 degrees Celsius, hatch, discard the dead chicken embryo within 48 hours, collect chicken embryo dead in 48～144 hours, gather allantoic fluid and amniotic fluid, separation obtains a strain duck viral hepatitis virus, this virus can be used as and prepare duck virus hepatitis inactivated vaccine after deactivation, can also further utilize the duck virus hepatitis inactivated vaccine preparing to prepare the antibody of being combined with this vaccine specific, this vaccine and antibody all can be used for treatment and the prevention of duck viral hepatitis.
For this reason, in one aspect of the invention, the present invention proposes duck viral hepatitis virus, this virus, with preserving number CGMCC No.7804, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center.According to the embodiment of the present invention; duck viral hepatitis virus is to separate and obtain the liver of the duck from suffering from duck viral hepatitis disease; the duck viral hepatitis virus strain toxicity that its separation obtains is good; there is good immunogenicity; can be used as inactivated vaccine and produce strain and check seed culture of viruses; according to one embodiment of the invention; utilize this duck viral hepatitis virus production duck virus hepatitis inactivated vaccine; effectively improve the immunogenicity of duck virus hepatitis inactivated vaccine, thereby further improve duck virus hepatitis inactivated vaccine security and immune period.
In another aspect of this invention; the present invention proposes above-mentioned duck viral hepatitis virus in the purposes of preparing in duck virus hepatitis inactivated vaccine; in one embodiment of the invention; according to duck viral hepatitis virus cross immunity, protection test proves, this duck viral hepatitis virus has good cross immunity provide protection.Therefore utilize this duck viral hepatitis virus production duck virus hepatitis inactivated vaccine, can effectively improve the immunogenicity of duck virus hepatitis inactivated vaccine, thereby further improve duck virus hepatitis inactivated vaccine immune period and immune effect.
The present invention proposes a kind of duck virus hepatitis inactivated vaccine more on the one hand of the present invention, according to the embodiment of the present invention, this duck virus hepatitis inactivated vaccine comprises the duck viral hepatitis virus of deactivation.According to the embodiment of the present invention, the immune duration of this duck virus hepatitis inactivated vaccine at least can maintain 4 months.According to the embodiment of the present invention, this duck virus hepatitis inactivated vaccine can be preserved 12 months under 2～8 degrees Celsius, and therefore this vaccine has satisfactory stability.
According to the embodiment of the present invention, above-mentioned duck virus hepatitis inactivated vaccine is inactivated, the inactivator kind that can adopt is also not particularly limited, as long as can effectively destroy the nucleic acid construct of duck viral hepatitis virus and make protein denaturation, but do not destroy antigenicity and the blood clotting of duck viral hepatitis virus.According to one embodiment of the invention, the inactivator adopting is formaldehyde, and formaldehyde can effectively destroy the DNA structure of duck viral hepatitis virus, and then reaches the object of deactivation duck viral hepatitis virus.According to the embodiment of the present invention, the inactivator formaldehyde final concentration adopting is also not particularly limited, as long as effectively deactivation duck viral hepatitis is viral.Described duck viral hepatitis virus is carried out to deactivation can be comprised: under 37 degrees Celsius, utilizing final concentration is that 0.1%～0.3% formaldehyde carries out inactivation treatment 48 hours to duck viral hepatitis virus-culturing fluid.Can effectively carry out deactivation to duck viral hepatitis virus thus.
In still another aspect of the invention, the present invention proposes a kind of method of preparing duck virus hepatitis inactivated vaccine, the method can comprise: duck viral hepatitis virus recited above is carried out to deactivation, to obtain the duck viral hepatitis virus through deactivation; Utilize through the duck viral hepatitis virus of deactivation and prepare water; And be that 1:3 carries out mixing and emulsifying stirring by water and oil phase according to volume ratio, to obtain this duck viral hepatitis viral inactivation vaccine.Thus, effectively improve the immunogenicity of duck virus hepatitis inactivated vaccine, thereby further improve duck virus hepatitis inactivated vaccine immune period and immune effect.
According to one embodiment of present invention, before duck viral hepatitis virus is carried out to deactivation, can obtain duck viral hepatitis virus-culturing fluid according to the following step: first, duck viral hepatitis virus inoculation, in nonimmune chicken embryo, and is cultivated under 35.5～37.5 degrees Celsius; Secondly, collect the chick embryo allantoic liquid obtaining, to obtain duck viral hepatitis virus antigen.According to the embodiment of the present invention, nonimmune chicken embryo is instar chicken embryo on the 11st～13, can effectively cultivate duck viral hepatitis virus.According to one embodiment of the invention, before nonimmune chicken embryo, first duck viral hepatitis virus is carried out rarely at duck viral hepatitis virus inoculation, after inoculation, at 37 ℃, cultivate, discard dead chicken embryo in 48 hours, collect 48～144 hours dead chicken embryos.Thereby be conducive to obtain the chick embryo allantoic liquid that malicious valency is high, and then improve the immune protective efficiency of preparing duck virus hepatitis inactivated vaccine.
According to one embodiment of the invention, duck viral hepatitis virus is carried out the condition of deactivation and is not particularly limited, according to a particular embodiment of the invention, can adopt formaldehyde to carry out deactivation to duck viral hepatitis virus.The inactivator formaldehyde final concentration adopting is also not particularly limited, as long as effectively deactivation duck viral hepatitis is viral.The final concentration of the formaldehyde adopting according to a particular embodiment of the invention, is 0.1%～0.3%.According to the embodiment of the present invention, the time of inactivator deactivation and temperature are also not particularly limited, as long as can effectively destroy the nucleic acid construct of duck viral hepatitis virus.According to a particular embodiment of the invention, utilizing final concentration is that 0.1%～0.3% formaldehyde carries out inactivation treatment 48 hours to duck viral hepatitis virus-culturing fluid, thus can be by the fully deactivation of duck viral hepatitis virus.
According to one embodiment of present invention, in the above-mentioned method of preparing duck virus hepatitis inactivated vaccine, can utilize following method to prepare water: the solution that contains deactivation duck viral hepatitis virus is mixed with tween-80.
According to a particular embodiment of the invention, the proportioning that the solution that contains deactivation duck viral hepatitis virus and tween-80 mix is also not particularly limited, and according to concrete example of the present invention, can mix according to the volume ratio of 96:4.Prepare water by adopting said ratio and concentration.
According to one embodiment of present invention, in the above-mentioned method of preparing duck virus hepatitis inactivated vaccine, can utilize following method to prepare oil phase: injection white oil MARCOL52, Si Ben-80 and aluminum stearate to be mixed, to obtain oil phase.According to a particular embodiment of the invention; the quality that injection white oil MARCOL52, Si Ben-80 and aluminum stearate can adopt is 94:6:2; the immunogenicity of duck virus hepatitis inactivated vaccine be can improve thus, thereby duck virus hepatitis inactivated vaccine immune period and immune protective effect further improved.
Utilize thus aforesaid method can effectively prepare immune period length and preferably duck virus hepatitis inactivated vaccine of immune protective effect.
In one side more of the present invention, the present invention proposes a kind of antibody.According to a particular embodiment of the invention, this antibodies specific is identified foregoing duck virus hepatitis inactivated vaccine.Utilize thus this antibody can be effective to treatment and preventing duck virus hepatitis.
Exempt from view of the domestic duck viral hepatitis having gone on the market is before high antibody exist unstable, the duck embryo of tiring can some homology disease of vertical transmission, duck idioplasm amount, the yellow antibody producing technique of the duck egg hidden danger, the antibody titer that the exist problem in measuring.In conjunction with above-mentioned practical situation, the present inventor adopts the method for immunization laying hen to produce the yellow antibody of the anti-duck viral hepatitis high-immunity egg of chicken by trial, be surprised to find that, the egg yolk antibody that adopts the method to produce, can use AGP method to carry out the inspection of antibody effect, and assay is reproducible, promptly and accurately, efficiently solve the problems referred to above that exist in production practice, and the antibody that production obtains can be effective to prevention and treatment duck viral hepatitis.
For this reason, in still another aspect of the invention, the present invention proposes a kind of method of stating antibody of preparing, according to one embodiment of present invention, the method comprises: the egg of giving birth to through foregoing duck virus hepatitis inactivated vaccine immunity chicken is provided; Separation obtains the yolk of egg, and adds the physiological saline of 3 times of volumes, to obtain egg yolk liquid; To in egg yolk liquid, add isopyknic trichloromethane also fully to mix, leave standstill 60 minutes, to obtain suspension; Suspension is carried out centrifugal and collects supernatant liquor; And be 0.05% formaldehyde solution by adding final concentration in supernatant liquor, to obtain this antibody.The method, by immunization laying hen, has extracted immunoglobulin (Ig) from yolk, therefore utilizes aforesaid method of the present invention can effectively utilize allos animal chicken to prepare the antibody of specific recognition duck virus hepatitis inactivated vaccine.The antibody simultaneously preparing is greater than 32 times for tiring, be qualified height and exempt from duck virus hepatitis yolk antibody.
According to a particular embodiment of the invention, in aforesaid method, be healthy chicken flock for immune chicken group, be qualified chicken embryo through immune chicken embryo.According to a particular embodiment of the invention, yolk is mixed according to the volume ratio of 1:3 with physiological saline, potent antibodies in yolk (livitin) fully can be dissolved in normal saline solution thus, can make thus the lipid material in yolk fully separate with aqueous substance).According to a particular embodiment of the invention, egg yolk liquid and trichloromethane are fully mixed according to the volume ratio of 1:1, and at room temperature leave standstill 60 minutes, and the lipid material in egg yolk liquid and water-soluble substances can be realized to layering thus, be convenient to abundant separation and extraction yolk antibody.Specifically, by adding trichloromethane can effectively extract fat, reducing the unnecessary material in antibody, static 60 minutes is exactly that lipid material is fully separated with aqueous substance.
According to one embodiment of present invention, the above-mentioned centrifugal rotating speed with 3500～4000rpm completes for centrifugal 30 minutes.Can remove thus lipid material, under this rotating speed, the lipid material little density aqueous substance large with density effectively can be separated.According to a particular embodiment of the invention, by adding the appropriate formaldehyde microorganism that effectively deactivation may be polluted or directly be propagated.Can further improve thus purity and the reliability of antibody, effectively solve the danger of this animal vertical transmission disease and the high allergenicity of serum, reduce production cost and use cost simultaneously.Utilize thus the antibody that aforesaid method prepares can be effective to the prevention of duck viral hepatitis and treat anti-.
Of the present invention more on the one hand, the present invention proposes a kind of being used for the treatment of or the medicine of preventing duck virus hepatitis.According to a particular embodiment of the invention, this pharmaceutical pack is containing antibody recited above.Can further improve thus that this medicine is used for the treatment of or preventing duck virus hepatitis specificity and drug effect.
Additional aspect of the present invention and advantage in the following description part provide, and part will become obviously from the following description, or recognize by practice of the present invention.
Embodiment
Below with reference to specific embodiment, present invention is described, it should be noted that, these embodiment are only descriptive, and do not limit the present invention in any way.
Embodiment 1 duck viral hepatitis virus
1, virus separates
In Shandong Mou Ya factory morbidity duck, gather its liver, rinse with sterile saline, discard more than mucous membrane, use sterilizing utensil to be shredded, grind and smash to pieces, add 2 times of sterile salines, it is mixed with tissue, tentatively filter with 100 order copper gauzes, 3500rpm centrifuging and taking supernatant, is finally used the degerming of 0.22um filter membrane again.By viral extracting solution inoculation 11～13 age in days SPF chick embryo allantoic cavities, hatch for 37 ℃, discard the dead chicken embryo within 48 hours, collect chicken embryo dead in 48～144 hours, gather allantoic fluid and amniotic fluid, separate and obtain duck viral hepatitis virus thus.
2, identify
Get the case of the doubtful viral duck hepatitis of Shandong generation, after tissue homogenate, further by the membrane filtration of 0.22 micron, filtrate inoculation SPF chicken embryo, results chick embryo allantoic liquid, obtains a strain virus, through being accredited as viral hepatitis virus, according to morbidity place, called after viral hepatitis virus (SH strain).This strain has been deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC) preservation on June 26th, 2013, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Classification And Nomenclature is duck viral hepatitis virus, and preserving number is CGMCC No.7804.
Embodiment 2 prepares duck virus hepatitis inactivated vaccine
Adopt the allantoic fluid for preparing of embodiment 1, by the formaldehyde solution that wherein adds 0.1%～0.3%, and airtight deactivation 48 hours at 37 ℃.After deactivation completes, preparation vaccine oil phase part, its ratio is 94 parts of white oils, these 806 parts, department, 2 parts of aluminum stearates, for subsequent use after heat sterilization; Preparation vaccine aqueous portion, its ratio is 96 parts, deactivation venom, 4 parts of tween 80s, the fully effect that makes it is dissolved completely.When emulsification, water, oil phase ratio are 1:3, and water oil mixt is added in mulser, and emulsification is prepared into duck virus hepatitis inactivated vaccine.
Embodiment 3 antibody preparation and application
1, prepare duck virus hepatitis inactivated vaccine antigen according to the method for embodiment 2, for subsequent use.
2, fundamental immunity: will treat that immune chicken injects the duck virus hepatitis inactivated vaccine antigen of above-mentioned preparation, be only specially neck subcutaneous injection 0.5ml/.
3, reinforced immunological: until fundamental immunity after 2 weeks, then carry out immunity for the second time, be only specially neck subcutaneous injection 1.0ml/; Interval 2 Zhou Houzai carry out reinforced immunological for the third time, are only specially neck subcutaneous injection 2.0ml/; At interval of 8 weeks, do primary reinforcement immunity, each neck subcutaneous injection 2.0ml/ only later.
4, plant egg collection: after reinforced immunological 7 days for the second time start, and get height exempt from egg yolk every three days, utilize agar diffusion (AGP) method to measure anti-duck viral hepatitis specific antibody titres.After yolk and physiological saline dilute according to 1:3 (V/V), add equal-volume trichloromethane extraction and carry, can start to collect above egg if supernatant liquor agar expansion antibody titer reaches 1:64, for the preparation of chicken viral hepatitis yolk antibody.5, antibody preparation: choose size evenly, appearance is clean without dirt, flawless egg, is soaked in 0.5% bromogeramine thimerosal 5 minutes,, more than water stain, beat eggs and fully remove egg white, blastodisc and frenulum in wiping outside, collects yolk.The sterile saline that adds 3 times of volumes in yolk, stirs, and then in egg yolk liquid, adds and the isopyknic trichloromethane of egg yolk liquid, fully concussion mixes, room temperature effect 60 minutes, with 3500～4000rpm centrifugal 30 minutes, collects supernatant liquid for subsequent use.
6, the pure property of antibody processing: add formaldehyde solution in the supernatant liquor separating, making it ultimate density is 0.05%, stir, 37 ℃ are incubated 8～12 hours, use again afterwards the degerming of 0.22um filtering with microporous membrane, aseptic subpackaged in suitable sterilising vessel, preparation becomes the high antibody finished product of exempting from of duck viral hepatitis.
7, inspection after construction
Steriling test, extracts the high antibody of exempting from of duck viral hepatitis, carries out steriling test with reference to " veterinary drug allusion quotation " (2010 editions, three) method, and goods are answered asepsis growth.
Physical behavior this product is micro-yellow or flaxen transparent liquid.Be long placed in rear bottle and have a little tiny white precipitate in an end.PH value 5.5～6.5.
5 of 18-22g healthy mices for safety verification, each subcutaneous injection this product 0.5ml, 3 10 of age in days susceptible ducklings, each subcutaneous injection this product 3.0ml/ is only.Observe 10 days, mouse and duckling should be all good for and be lived as qualified.
AGP experiment: it is qualified above that antibody titer is measured >=64 times.
Efficacy test: with the inspection of duck effect; 3～5 30 of age in days susceptible ducklings, are divided into three groups at random.First group is normal healthy controls group, does not inject any medicine, isolated rearing.Second group and the 3rd group every the strong malicious SCG99 strain enteron aisle pathological material of disease poison of duckling subcutaneous vaccination duck viral hepatitis, make it viral level and be not less than 100LD
50/ 0.1ml, 0.1ml/ only.In attacking malicious latter 24 hours respectively to second group of subcutaneous injection this product 2.0ml, the 3rd group of subcutaneous injection physiological saline 2.0ml.Observe every group of duckling morbidity, death condition to the 10 days.
Result is judged: first group is normal healthy controls group, and the whole ducklings of duration of test should be healthy anosis.The 3rd group for attacking malicious control group, and Ying Yu attacks morbidity in 3～7 days after poison, starts deadly after 96 hours, and within 10 days, planted agent is dead more than 8.Second group is early infection treatment group, and injection this product after 48 hours, attacks poison latter 3 days, and experiment duckling starts morbidity, and sickness rate is higher than 20%, and while end to experiment, duckling should survive and 8 only reach above, and it is qualified that these goods are judged to.
8, can be found out by above experimental result, the anti-duck viral hepatitis yolk antibody of chicken has good preventive and therapeutic action for Duck Hepatitis Virus epidemic disease.In this embodiment, used the detection method of agar diffusion (AGP) to carry out the detection that duck viral hepatitis yolk antibody is tired, detected result is accurate rapidly, reproducible.The anti-duck viral hepatitis egg yolk antibody of producing by the inventive method, can solve the danger of this animal vertical transmission disease and the high allergenicity of serum, has reduced production cost and use cost simultaneously.It is certain that height is exempted from antibody preventing viral hepatitis effect.
In the description of this specification sheets, the description of reference term " embodiment ", " some embodiment ", " example ", " concrete example " or " some examples " etc. means to be contained at least one embodiment of the present invention or example in conjunction with specific features, structure, material or the feature of this embodiment or example description.In this manual, the schematic statement of above-mentioned term is not necessarily referred to identical embodiment or example.And specific features, structure, material or the feature of description can be with suitable mode combination in any one or more embodiment or example.
Although illustrated and described embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change above-described embodiment within the scope of the invention in the situation that not departing from principle of the present invention and aim, modification, replacement and modification.
Claims (13)
1. a duck viral hepatitis virus, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center with preserving number CGMCC No.7804.
2. duck viral hepatitis virus claimed in claim 1 is in the purposes of preparing in duck virus hepatitis inactivated vaccine.
3. a duck virus hepatitis inactivated vaccine, is characterized in that, comprises:
The duck viral hepatitis virus claimed in claim 1 of deactivation.
4. duck virus hepatitis inactivated vaccine according to claim 3, is characterized in that, described duck viral hepatitis virus is by formalin-inactivated.
5. a method of preparing duck virus hepatitis inactivated vaccine, is characterized in that, comprising:
Duck viral hepatitis virus claimed in claim 1 is carried out to deactivation, to obtain the duck viral hepatitis virus through deactivation;
Utilize the described duck viral hepatitis virus through deactivation to prepare water; And
Be that 1:3 carries out mixing and emulsifying stirring by described water and oil phase according to volume ratio, to obtain described duck viral hepatitis viral inactivation vaccine.
6. method according to claim 5, is characterized in that, before described duck viral hepatitis virus is carried out to deactivation, comprises according to the following step and obtains duck viral hepatitis virus-culturing fluid:
Described duck viral hepatitis virus inoculation, in nonimmune chicken embryo, and is cultivated under 35.5～37.5 degrees Celsius;
Collect the chick embryo allantoic liquid obtaining, to obtain described duck viral hepatitis virus antigen.
7. method according to claim 5, is characterized in that, described duck viral hepatitis virus is carried out to deactivation and comprise:
Under 37 degrees Celsius, utilizing final concentration is that 0.1%～0.3% formaldehyde carries out inactivation treatment 48 hours to duck viral hepatitis virus-culturing fluid.
8. method according to claim 5, is characterized in that, the described water of preparing comprises:
The solution that contains deactivation duck viral hepatitis virus is mixed according to the volume ratio of 96:4 with tween-80, to obtain described water.
9. method according to claim 5, is characterized in that, the described oil phase of preparing comprises:
Injection white oil MARCOL52, Si Ben-80 and aluminum stearate are mixed according to the mass ratio of 94:6:2, to obtain described oil phase.
10. an antibody, is characterized in that, described duck viral hepatitis antibodies specific is identified the duck virus hepatitis inactivated vaccine described in claim 3 or 4.
The method of 11. 1 kinds of Dispersal risks, is characterized in that, comprising:
The egg of giving birth to through the duck virus hepatitis inactivated vaccine immunity chicken described in claim 3 or 4 is provided;
Separation obtains the egg yolk of described egg, and adds the physiological saline of 3 times of volumes, to obtain egg yolk liquid;
To in described egg yolk liquid, add isopyknic trichloromethane also fully to mix, leave standstill 60 minutes, to obtain suspension;
Described suspension is carried out centrifugal and collects supernatant liquor; And
Be 0.05% formaldehyde solution by adding final concentration in described supernatant liquor, to obtain described duck viral hepatitis antibody.
12. methods according to claim 11, is characterized in that, the described centrifugal rotating speed with 3500～4000rpm completes for centrifugal 30 minutes.
13. 1 kinds are used for the treatment of or the medicine of preventing duck virus hepatitis, it is characterized in that, comprise antibody claimed in claim 10.
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| CN1927884A (en) * | 2005-09-09 | 2007-03-14 | 重庆永健生物技术有限责任公司 | Preparation method of heterogenous yolk antibody for preventing and curing duck viral hepatitis |
| CN101607994A (en) * | 2008-06-18 | 2009-12-23 | 洛阳普莱柯生物工程有限公司 | A kind of preparation method of duck viral hepatitis refine yolk antibody |
| CN101612397A (en) * | 2009-07-17 | 2009-12-30 | 齐鲁动物保健品有限公司 | A kind of duck virus hepatitis inactivated vaccine and preparation method thereof |
| CN102078609A (en) * | 2010-12-23 | 2011-06-01 | 天津康莱森生物科技集团有限公司 | Method for preparing duck virus hepatitis refined heterogenous yolk antibody |
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- 2013-08-14 CN CN201310354525.0A patent/CN103834619A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1927884A (en) * | 2005-09-09 | 2007-03-14 | 重庆永健生物技术有限责任公司 | Preparation method of heterogenous yolk antibody for preventing and curing duck viral hepatitis |
| CN101607994A (en) * | 2008-06-18 | 2009-12-23 | 洛阳普莱柯生物工程有限公司 | A kind of preparation method of duck viral hepatitis refine yolk antibody |
| CN101612397A (en) * | 2009-07-17 | 2009-12-30 | 齐鲁动物保健品有限公司 | A kind of duck virus hepatitis inactivated vaccine and preparation method thereof |
| CN102078609A (en) * | 2010-12-23 | 2011-06-01 | 天津康莱森生物科技集团有限公司 | Method for preparing duck virus hepatitis refined heterogenous yolk antibody |
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