CN1958069A - Triple high immunity immunoglobulin for anti swine pest, aujeszkys disease of pig, and parvovirus of pig - Google Patents
Triple high immunity immunoglobulin for anti swine pest, aujeszkys disease of pig, and parvovirus of pig Download PDFInfo
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Abstract
An immunoglobulin for preventing the cholera, pseudorabies and parvovirus of hog is prepared from their immunogens through proportional mixing them together and proportionally mixing it with complete Frennd's adjuvant. It features that the glycoside-peptide injection is used as its immunopotentiator.
Description
Technical field
The invention belongs to biological technical field, be specifically related to anti-swine fever, the porcine pseudorabies of three kinds of immunogen preparing of a kind of employing, the immunoglobulin of pig parvoviral, international Patent classificating number is: A61K39/395.
Background technology
Swine fever (Classical swine fever CSF, Hog chorera HC) is a kind of crushing infectious disease of pig, is hog cholera again in one's early years, is a kind of deadly infectious disease that is caused by swine fever virus.Since tennessee,USA reported first in 1810, be global popular.In recent years, removing the North America, outside Oceania and European a few countries are on the shelf, remain the biggest threat of pig industry, is the most important infectious disease of present pig.The International Health Regulation of International Office of Epizootics (OIE) is classified swine fever as one of deadly infectious disease of 16 kinds of category-A researchs; Chronic through out-of-date, how with other antibacterial secondary infection, make state of an illness aggravation.
Swine fever is that (Hog cholera virus HCV) causes by the swine fever virus of flaviviridae (Flaviviridae) pestivirus (Pestivirus).It is alternate that swine fever shows as heating, anorexia, diarrhoea and constipation not immune swinery, typical clinical symptoms such as depletion, death.In the clinical manifestation of immunological incompetence swinery, from asymptomatic, subclinical infection is dead to typical case's morbidity, differs greatly.Can present complicated breeding dysfunction syndrome shapes such as stillborn fetus, miscarriage, weak son, piglet survival ratio be low in-pig.
The pseudorabies of pig is one of deadly infectious disease of harm pig industry, is the multiple animal that caused by Pseudorabies virus and a kind of acute infectious disease of birds.In an area that pseudorabies did not take place, reach more than 90% the death of pig farm piglet that has even one-tenth nest during outburst to the preceding piglet mortality rate of wean from birth.The pseudorabies M ﹠ M of growing and fattening pigs is also all very high, but is usually ignored by people.
On taxonomy, (Pseudorabies virus PRV) is the DNA viruses that belongs to herpetoviridae (Herpesviridae) Varicellavirus (Varicellavirus) to Pseudorabies virus.Virus is spherical in shape, and cyst membrane is arranged, can be in multiple mammalian cell internal breeding, and produce intranuclear inclusion.This virus often is present in the marrowbrain tissue, and sick pig also contains virus at fb dur among its nose liquid, saliva, milk, vaginal secretions, blood and the organa parenchymatosum.
Porcine pseudorabies has 4 big clinical manifestations: (1) in-pig is miscarried, stillborn fetus, mummy tire, wherein to produce stillborn fetus; (2) newborn piglet death, 15 ages in days reach 100% with interior piglet mortality rate.Normal performance sudden onset, body temperature rises to more than 41 ℃, anorexia (breast), more than, the lethargy of vomitting, have loose bowels, shakes, moves inharmonious, uses antibiotic, and sulfa drugs is failed to respond to any medical treatment.Septicemia and nervous symptoms are arranged, especially significantly straight-forward phenomenon be an ear forward, an ear is backwards; (3) the ablactational baby pig sickness rate is about 40%, and mortality rate about 20% mainly shows as Respiratory symptoms, is dyspnea, cough, rhinorrhea etc.; Also there is the part pig nervous symptoms, diarrhoea and vomiting etc. to occur; The wiping distinctive symptom of other domestic animals often of itching, and pig does not have, but the present ill baby pig disease Cheng Qian phase present and wipe itch increasing, show that this virus has the enhanced sign of virulence in popular process; (4) sow performance infertility is returned the feelings rate and is reached 90%, and breeding all unmatches for several times.
It is that pig parvoviral (Porcine parvovirus PPV) by Parvoviridae (Parvovirioae) parvovirus belongs to (Parvovirus) causes one of main pathogen of sow breeding difficulty that pig parvoviral infects.Be with embryo and fetal infection and death and parent itself does not show a broad sow breeding difficulty sexually transmitted disease of symptom.In swinery all over the world, this virus ubiquity, it is popular to be endemicity on most of pig farms.
Infect the PPV initial stage, food, asthenia, vomiting or diarrhoea may occur subtracting, slightly be difficult for being discovered with symptom because of the time is of short duration.Therefore, the pig of infection PPV all shows as inferior clinical symptom usually.Its cardinal symptom is a Disease in Infants breeding anergy.The infection oestrus of sow is undesired, joins infertile for a long time.It is also variant because of different its clinical responses of gestational age that in-pig infects PPV, and the gestation initial stage infects, and the cause death fetus is absorbed, and farrowing quantity is reduced; Miscarriage and normal producing can appear in the intermediary and later stages infection, and the normal piglets of output accounts for 70%, and unusual accounts for 30%, and young unusually is stillborn fetus, weak tire and mummy tire.The unusual piglet of survival can not stand up, weak, very fast death.The sow that breeding difficulty takes place except that miscarriage, stillborn fetus, mummy appear and the phenomenon such as infertile of oestrusing, most of no tangible clinical symptoms, indivedual sows have fervescence, the rear quarters motion is dumb or paralyse, arthroncus or body surface have phenomenons such as circular swelling.
For the prevention of these three kinds of diseases, the policy that China takes at present mainly is to prevent with vaccine.But because China's present feeding manner great majority are still in the majority with the person of raising scattered of peasant household, many peasant households, happen occasionally owing to omit the phenomenon of immunity in many areas to reasons such as the understanding deficiency of vaccine immunity and systems, and these three kinds of diseases also become worse.In addition, from these three kinds of diseases itself since its popular comparatively rapidly and the infection rate in swinery higher, so its M ﹠ M is also higher.Especially these three kinds of diseases all can cause the breeding difficulty of sow, and sow breeding difficulty disease is the pig industry development of restriction China, cause the important diseases of heavy economic losses.Swine fever, porcine pseudorabies, parvovirus infections not only can cause sow miscarriage, stillborn fetus, premature labor, few product, produce the mummy tire and produce weak young.Also can infect the piglet of output simultaneously, cause the piglet mortality.This class disease has the feature of mass-sending, and caused breeding potential descends has become in pig farm, family, especially the intensive pig production field comparatively common phenomena.Economic loss is very serious.
On the other hand, treatment for viral disease, also not as antibiotic to the unusual specific medicament of the antiviral property disease the specially good effect of antibacterial, after the animal morbidity, use antibiotics or immunomodulator, just, can not tackle the problem at its root in order to control the non-specific immunity of concurrent bacterial infection and raising animal body.And, after the swinery morbidity, caused the fact of sow miscarriage and piglet death, heal with medicine late this moment, because expended feeding cost, and do not obtain expecting the piglet of wishing, also have to eliminate a part of sow simultaneously at the sow phenolics.These three kinds of diseases are very huge to the loss that pig industry caused thus, have become to have of overall importance, critical problem in the Swine Production.
Summary of the invention
Goal of the invention of the present invention is to adopt three kinds of immunogen preparing to have the specificity height, and is with strong points, treatment that curative effect is certain or prevention swine fever, porcine pseudorabies, the anti-swine fever of pig parvoviral, porcine pseudorabies, pig parvoviral triad high immune immunoglobulin.
The immunogen that the present invention adopts adds complete Freund adjuvant after concentrated and is prepared from getting cell culture, wherein swine fever virus is cultivated with bull testis primary cell or PHK-21 passage cell, porcine pseudorabies virus is cultivated with porcine kidney cell (PK-15) or bovine kidney cells (MDBK), pig parvoviral former generation of Ren sus domestica or subculture cell culture; Concentrate tiring of these three kinds of viruses of back and reach 10
6～8, with these three kinds of immunogens with volume ratio (1～3): (1～3): add Freund adjuvant at 1: 1 as the immunogen immune animal according to volume ratio after the mixed of (1～3).(denomination of invention is the Heterosides peptide injection that uses the Jilin whale to resemble animal pharmaceutcal corporation, Ltd simultaneously to be developed into: a kind of Heterosides peptide injection that improves the animal immune function, number of patent application is 2,005 10016626.2) as immunostimulant, 5 milliliters/, to improve antibody titer.
Immune programme for children
Immune animal of the present invention is used through the pig of strict quarantine, horse (mule, donkey) as immune animal.
Immunization route adopts Intradermal, subcutaneous or acupuncture point, muscle multi-point injection.Fundamental immunity dosage is every pig 3ml, horse (mule, donkey) 5ml.The immunization ways pig is subcutaneous or the muscle multi-point injection, and horse (mule, donkey) is used Intradermal, subcutaneous or acupuncture point multi-point injection.Carried out the booster immunization first time in 18～21 days at interval, immunizing dose is pig 8ml, horse (mule, donkey) 10～12ml; Booster immunization was carried out once in 18～21 days in later every interval, behind 2～4 booster immunizations, detect antibody titer, it is qualified that antibody titer 〉=2048 o'clock are considered as, do not reach this standard as antibody titer, continue booster immunization once, then to the blood sampling of horse (mule, donkey) jugular vein or to immune swine carotid artery or the direct blood-letting of heart.
Antibody titer detects
Use the ELISA method that detects these three kinds of diseases to detect serum antibody titer respectively.Detect antigen and use cell culture through virus spissated and the process respective handling, second antibody adopts anti-pig of corresponding rabbit (mouse-anti pig) or the anti-horse of rabbit (mouse-anti horse), sets up the contrast of positive and negative serum simultaneously.The serum antibody titer eligible can be taken a blood sample or dispatch from the factory as qualified products.
Blood sampling and serum extract
At first drench fully with the container inner wall of 1%～5% aseptic sodium chloride solution with blood sampling.Each container should be indicated animal number, last booster immunization date and blood sampling date.Behind the sterile blood sampling with the blood gathered 20 ℃～25 ℃ of placements, treat that it all solidifies and when having serum to separate out, adds the ram of sterilization, after 48～72 hours, extracts serum with sterile manner.Press 0.5% of serum total amount simultaneously and add chloroform, 0.0035% adding thimerosal, the sealing of jumping a queue fully shakes up, and is positioned over 2 ℃～15 ℃ dark cold places, the precipitation clarification.Simultaneously every bottle of serum should post animal number, serum amount and separate the date.
Blood plasma extracts
In the sterilization vial that fills 5% sterilization liquor sodii citratis, the ratio of blood and liquor sodii citratis is 9: 1 with blood collecting; Through 48～72 hours precipitations, to extract supernatant, and add chloroform, 0.0035% adding thimerosal by 0.5% of total amount, the sealing of jumping a queue fully shakes up.Every bottle of blood plasma should be made itemized record.
Serum (blood plasma) is refining
Will be mixed in the same sterilization container by purified serum (blood plasma), 2 times of amounts of pressing serum (blood plasma) total amount add distilled water, stir; Regulate pH to 3.0 with 2mol/L hydrochloric acid, when adding hydrochloric acid adjusting pH, slowly add, the limit edged stirs the limit and measures, and prevents that the local pH peracid from exerting an influence to the activity of immunoglobulin.Placed 60 minutes, the sodium hydroxide solution of reuse 2mol/L is regulated pH to 3.2～3.4.Amount according to every 1ml 6～9 units adds pepsin then, and adds toluene by 0.2% of total amount, digests 2～24 hours under 29 ℃～31 ℃ conditions, will often stir between the period of digestion.
The solution that has digested is added ammonium sulfate according to 15% of total amount, fully after the dissolving, regulate pH to 4.5～4.7,60 ℃ water-bath 60 minutes with the sodium hydroxide solution of 2mol/L.When treating that solution is cooled to below 43 ℃, filter, collect filtrate, remove precipitation with canvas.The filtrate of collecting is measured, and be put in the container of same sterilization, regulate pH to 7.1～7.3, add ammonium sulfate, after stirring, filter the collecting precipitation thing with the canvas of sterilizing according to 20% of total amount with the sodium hydroxide solution of 2mol/L.Then precipitation is carried out weighing, put in the sterile chamber, and in precipitate, add 5～6 times distilled water, the dissolution precipitation thing, treat that precipitate fully dissolves after, alum solution according to total amount adding 10%, make the Alumen total content reach 1%, reuse 2mol/L sodium hydroxide solution is regulated pH to 7.7～7.8, leaves standstill and makes its precipitation in 2～4 hours, filter with aseptic canvas then, collect filtrate.
The filtrate of collecting is positioned in the same sterile chamber, add ammonium sulfate by 38% of total amount, fully stirring the back filters with aseptic canvas, abandon the supernatant collecting precipitation, precipitation is pressed dry back bag filter bag distribution packaging, every bag of about 200～250g adds a little chloroform, tightens to be positioned in the gutter channel dialysis behind the bag filter 48～72 hours.Serum in the collecting bag adds chloroform, 0.85% by 0.5% of total amount and adds sodium chloride then, and 0.0035% adds thimerosal.Fully the 0.22um membrane filtration is used in the dissolving back under aseptic condition, is positioned over 5 ℃～15 ℃ and staticly settles.The serum that takes a morsel then carries out antibody titer and detects, and its protection effectiveness is measured.
According to " People's Republic of China's veterinary biologics rules " that The Ministry of Agriculture of the People's Republic of China, MOA published in 2000 the serum of producing is carried out steriling test.
High immune immunoglobulin is to swine fever and porcine pseudorabies immune efficacy determination test
It is 4 groups that 20 of 15kg left and right sides susceptible piglets are divided into, 5 every group.First group of intramuscular injection 10
6～8The swine fever virus virulent strain of tiring, injection detects the high immune immunoglobulin 10ml of antibody titer 〉=2048 after 8～12 hours through ELISA, inject once again after 48 or 72 hours, dosage still is 10ml, observed for 2 weeks continuously, after the off-test, the servant does pathological anatomy and histopathologic examination extremely with test pig.During this period, the typical clinical symptom occurs person and pathological anatomy and histopathologic examination swine fever typical case pathological change person occurs and is judged to infection, clinical symptoms do not occur and pathology variation person is judged to protection.Second group is matched group, and processing method is identical with first group, just uses the physiologic saline for substitute high immune immunoglobulin behind the viral infection.The 3rd group of intramuscular injection 10
8The porcine pseudorabies virus virulent strain of tiring, injection is injected once after 48 hours through the high immune immunoglobulin 10ml of ELISA detection antibody titer 〉=2048 again after 48～72 hours, observes for 2 weeks continuously, after the off-test, the servant does pathological anatomy and histopathologic examination extremely with test pig.During this period, the typical clinical symptom occurs person and pathological anatomy and histopathologic examination porcine pseudorabies type pathological change person occurs and is judged to infection, clinical symptoms do not occur and pathology variation person is judged to protection.The 4th group is matched group, and processing method is identical with first group, just uses the physiologic saline for substitute high immune immunoglobulin behind the viral infection.
To swine fever and porcine pseudorabies immune efficacy measurement result
First group and the 3rd group be all protections clinically, just have a pig phenomenon of slight lymphadenitis to occur in first group, occur the interior capillary endothelial cell enlargement of slight lymph node.Second group of 3/5 clinical onset in the matched group, all the other two are positive through pathological examination.Petechia not of uniform size promptly appears in the wherein pathological change of a pig appearance " bird egg kidney " under the tunicle of kidney.The variation of lymph node periphery edema and diffuse hemorrhage appears in the other end pig.Be judged to morbidity.In the 4th group of matched group, tangible clinical symptoms appears in 4/5 clinical onset, and dead 3, fervescence appears in the other end, and appetite reduces, symptoms such as poor growth.It is hemorrhage that the other end does not have the swine diseases reason of clinical symptoms to cut open the enlargement of inspection discovery colic lymph nodes, and endocardium and kidney have petechia and typical inflammatory activity, are judged to morbidity.
High immune immunoglobulin is to pig parvoviral immune efficacy determination test
Three in-pigs are divided into one group, at the high immune immunoglobulin 50ml of injection in antenatal 5 days, simultaneously intramuscular injection 10 in antenatal 5 days through ELISA detection antibody titer 〉=2048
6～8The strong poison of the pig parvoviral of tiring carries out challenge test, observes continuously to 21 days puerperal, and the piglet of output does not fall ill and is judged to protection, and the piglet morbidity is judged to infection.Experiment is carried out pathological examination after finishing.With 2 in-pigs in contrast, do not inject high immune immunoglobulin, an injecting normal saline, the counteracting toxic substances method is the same, after the off-test, the farrowing pig is carried out pathological examination, log.
High immune immunoglobulin is to pig parvoviral immune efficacy measurement result
Three test sow 32 of piglets of common property (9,12,11), wherein because of dead 2 of other reasons, last 30 piglets do not see the piglet that clinical onset is arranged, fully protection.2 sow common property piglets 19 (10,9) of contrast are because dead 1 of other reasons, 12 of 18 piglet clinical onsets, and all death during puerperal 10 wherein has 9 nervous symptoms to occur.Last 5 slight clinical symptoms only occurs, find through pathological examination, and the pathology that all have weight not wait change.1 pig does not fall ill, and pathological examination does not see that pathology change yet.
The product packing
After suitably diluting, during as the finished product packing, antibody titer should exceed 20% than required standard with semi-finished product, even the antibody titer of time spent is 〉=2048 o'clock, the antibody titer of packing finished product answers 〉=2640.Using dosage is the 0.4ml/kg body weight.
The present invention has the following advantages and good effect:
1. the good high immune immunoglobulin of clinical application effect is treated these three kinds of viral diseases and is had high specificity, and curative effect is certain, the characteristics of short treating period, instant effect.Owing to hyper-immune serum prepares with these three kinds of virus immunity animals,, this this a pair of Swine Production of three kinds of diseases that virus causes, especially sow breeding difficulty prepared a species specific treatment preparation so being influenced earthshaking disease.
2. immunoglobulin half-life is short, does not have cumulative action in animal body, thus after with it these three kinds of diseases being prevented, treated the drug residue phenomenon can not appear, for consumer and export pork product and all have active operation significance.
3. result of the test shows, before farrowing 15～30 days, inject to sow with anti-swine fever, porcine pseudorabies, pig parvoviral triad high immune immunoglobulin, can effectively prevent the sow breeding difficulty that causes because of these three kinds of diseases, after giving the injection of morbidity sow with anti-swine fever, porcine pseudorabies, pig parvoviral triad high immune immunoglobulin simultaneously, can reach effective therapeutic purposes to its breeding difficulty.
4. safety uses pepsin that immunoglobulin is digested in the processing technique of immunoglobulin fortunately, the FC segment of antibody is cut away, make the molecular weight of immunoglobulin dwindle one times nearly, with the ammonium sulphate precipitation method albumin of blood weight is removed simultaneously, so just can not caused allergic reaction.
5. preservation and this product easy to use are light yellow transparent liquid, preserve for 4 ℃ and detect antibody titer in 1 year still more than 〉=2048, and be highly stable.In the use, a pin can prevent and treat three kinds of diseases.
The specific embodiment
Embodiment 1:
The immunogen that the present invention adopts adds complete Freund adjuvant after concentrated and is prepared from getting cell culture, wherein swine fever virus is cultivated with bull testis primary cell or PHK-21 passage cell, porcine pseudorabies virus is cultivated with porcine kidney cell (PK-15) or bovine kidney cells (MDBK), pig parvoviral former generation of Ren sus domestica or subculture cell culture; Concentrate tiring of these three kinds of viruses of back and reach 10
6～8, these three kinds of immunogens are added Freund adjuvant as the immunogen immune animal according to volume ratio 1: 1 after with 1: 2: 1.5 mixed of volume ratio.Use the Jilin whale to resemble Heterosides peptide injection that animal pharmaceutcal corporation, Ltd is developed into simultaneously as immunostimulant, 5 milliliters/, to improve antibody titer.
Preparation method of the present invention:
Immune programme for children
Immune animal of the present invention is used through the pig of strict quarantine, horse (mule, donkey) as immune animal.
Immunization route: adopt Intradermal, subcutaneous or acupuncture point, muscle multi-point injection.Fundamental immunity: every pig 3ml, horse (mule, donkey) 5ml.The immunization ways pig is subcutaneous or the muscle multi-point injection, and horse (mule, donkey) is used Intradermal, subcutaneous or acupuncture point multi-point injection.Booster immunization: carried out the booster immunization first time in 18～21 days at interval, immunizing dose is pig 8ml, horse (mule, donkey) 10～12ml.Booster immunization was carried out once in 18～21 days in later every interval, and behind 2～4 booster immunizations, antibody titer detects: use the ELISA method that detects these three kinds of diseases to detect serum antibody titer respectively.It is qualified that antibody titer 〉=2048 o'clock are considered as, and do not reach this standard as antibody titer, continues booster immunization once, then to the blood sampling of horse (mule, donkey) jugular vein or to immune swine carotid artery or the direct blood-letting of heart.
Blood sampling and serum extract
Blood sampling: behind the sterile blood sampling with the blood gathered at 20 ℃～25 ℃ of placements.Serum extracts: treat that it all solidifies and when having serum to separate out, adds the ram of sterilization, after 48～72 hours, extracts serum with sterile manner.Press 0.5% of serum total amount simultaneously and add chloroform, 0.0035% adding thimerosal, the sealing of jumping a queue fully shakes up, and is positioned over 2 ℃～15 ℃ dark cold places, the precipitation clarification.
Blood plasma extracts
In the sterilization vial that fills 5% sterilization liquor sodii citratis, the ratio of blood and liquor sodii citratis is 9: 1 with blood collecting; Through 48～72 hours precipitations, extract supernatant.
Serum (blood plasma) is refining
Will be mixed in the same sterilization container by purified serum (blood plasma), by 2 times of amount adding distilled water of total amount, regulate pH to 3.0 with 2mol/L hydrochloric acid, placed 60 minutes, the sodium hydroxide solution of reuse 2mol/L is regulated pH to 3.2～3.4.Amount according to every 1ml 6～9 units adds pepsin then, digests 2～24 hours under 29 ℃～31 ℃ conditions.
The solution that has digested is added ammonium sulfate according to 15% of total amount, regulate pH to 4.5～4.7,60 ℃ water-bath 60 minutes with the sodium hydroxide solution of 2mol/L.When treating that solution is cooled to below 43 ℃, filter, collect filtrate, remove precipitation.Metering also is put in the same container, regulates pH to 7.1～7.3 with the sodium hydroxide solution of 2mol/L, adds ammonium sulfate according to 20% of total amount, uses the canvas of sterilization to filter the collecting precipitation thing.Put into after the weighing in the container, add 5～6 times distilled water, behind the dissolution precipitation thing, according to the alum solution of total amount adding 10%, reuse 2mol/L sodium hydroxide solution is regulated pH to 7.7～7.8, leaves standstill and makes its precipitation in 2～4 hours, filters and also collects filtrate.
The filtrate of collecting is positioned in the same container, adds ammonium sulfate, stir the back and filter with aseptic canvas by 38% of total amount, abandon the supernatant collecting precipitation, precipitation is pressed dry the back use the bag filter bag distribution packaging, every bag of about 200～250g tightens to be positioned over behind the bag filter in the gutter channel and dialysed 48～72 hours.Serum in the collecting bag, fully the 0.22um membrane filtration is used in the dissolving back under aseptic condition, is positioned over 5 ℃～15 ℃ and staticly settles.
According to " People's Republic of China's veterinary biologics rules " that The Ministry of Agriculture of the People's Republic of China, MOA published in 2000 the serum of producing is carried out steriling test.
The product packing
After suitably diluting, during as the finished product packing, antibody titer should exceed 20% than required standard with semi-finished product, even the antibody titer of time spent is 〉=2048 o'clock, the antibody titer of packing finished product answers 〉=2640.Using dosage is the 0.4ml/kg body weight.
Embodiment 2:
Swine fever virus is cultivated with bull testis primary cell or PHK-21 passage cell, and porcine pseudorabies virus is cultivated with porcine kidney cell (PK-15) or bovine kidney cells (MDBK), pig parvoviral former generation of Ren sus domestica or subculture cell culture; Concentrate tiring of these three kinds of viruses of back and reach 10
6～8, these three kinds of immunogens are added Freund adjuvant as the immunogen immune animal according to volume ratio 1: 1 after with 1.5: 1: 1.5 mixed of volume ratio.Use the Jilin whale to resemble Heterosides peptide injection that animal pharmaceutcal corporation, Ltd is developed into simultaneously as immunostimulant, 5 milliliters/, to improve antibody titer.
Preparation method is identical with embodiment 1.
Claims (3)
1, anti-swine fever, porcine pseudorabies, pig parvoviral triad high immune immunoglobulin is characterized in that:
Immunogen adopts swine fever virus to cultivate with bull testis primary cell or PHK-21 passage cell, and porcine pseudorabies virus is cultivated with porcine kidney cell (PK-15) or bovine kidney cells (MDBK), pig parvoviral former generation of Ren sus domestica or subculture cell culture;
Immunological adjuvant: adopt Freund's complete adjuvant;
Immunostimulant: adopt the Jilin whale to resemble the Heterosides peptide injection that animal pharmaceutcal corporation, Ltd produces.
2, anti-swine fever according to claim 1, porcine pseudorabies, pig parvoviral triad high immune immunoglobulin is characterized in that:
With described three kinds of immunogens with volume ratio (1～3): (1～3): after the mixed of (1～3), with immunogen and Freund's complete adjuvant volume ratio according to 1: 1 mixed immunity animal;
The immunostimulant Heterosides peptide injection is 5 milliliters/.
3, anti-swine fever according to claim 1, porcine pseudorabies, pig parvoviral triad high immune immunoglobulin is characterized in that:
Described immunogen concentrates tiring of back three kinds of viruses and reaches 10
6～8
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101862452A (en) * | 2010-07-22 | 2010-10-20 | 山西芮城亚宝兽药有限公司 | Method for preparing swine multi-valent antiserum compound preparation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101862452A (en) * | 2010-07-22 | 2010-10-20 | 山西芮城亚宝兽药有限公司 | Method for preparing swine multi-valent antiserum compound preparation |
| CN101862452B (en) * | 2010-07-22 | 2012-06-20 | 山西芮城亚宝兽药有限公司 | Method for preparing swine multi-valent antiserum compound preparation |
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