CN104208680B - A kind of anti-pig wins sick freeze-dried mixed powder of card and preparation method thereof - Google Patents
A kind of anti-pig wins sick freeze-dried mixed powder of card and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of anti-pig and win sick freeze-dried mixed powder of card and preparation method thereof, this freeze-dried mixed powder includes the component of following parts by weight: anti-pig is rich blocks sick yolk antibody 94～97 parts, the sick transfer factor 1 of the rich card of anti-pig～2 parts, preservative 0.01～0.02 part, inactivator 0.2～0.3 part, selenium 0.05～0.1 part, vitamin C 0.5～1 part, polyvinylpyrrolidone 1～2 parts, glycine 0.5～1 part.The anti-pig of the present invention wins the freeze-dried mixed powder that card is sick, the sick transfer factor of card rich with anti-pig of rich for the anti-pig sick yolk antibody of card is compounded, and it is aided with multiple auxiliary materials, each component cooperates, coordinative role, its preventing and treating pig can be significantly improved and win the effect that card is sick, there is advantage with strong points, that room temperature lower holding time length, antibody titer high, biological activity is good, be a kind of potent biological preparation with wide application prospect.
Description
Technical field
The invention belongs to veterinary drug technical field, be specifically related to a kind of anti-pig and win the freeze-dried mixed powder that card is sick, also relate to one
Anti-pig wins the preparation method of the sick freeze-dried mixed powder of card.
Background technology
Pig-breeding occupies heavier share, the only Carnis Sus domestica of Henan Province's output in 2009 in China's aquaculture and just reaches 410.32
Ten thousand tons, it follows that China's of demand to(for) Carnis Sus domestica every year is the biggest.But, along with Carnis Sus domestica demand cumulative year after year and
, also there is increasing problem in the expansion of cultivation scale during pig-breeding, wherein pig wins card disease is a kind of acute infection
Property disease, causes huge economic loss to pig industry.
Pig bocavirus (Bocavirus) is under the jurisdiction of the bocavirus of Parvoviridae parvovirus subfamily and belongs to, this DNA
Virus is that China finds for nearest 2 years, has the clinical epidemiological features such as local dead rate frequently-occurring, high, highly infective, mainly
It is aimed at piglet, after weaned piglet, easily causes the diseases such as multisystemic exhaustion syndrome, transmissible gastroenteritis, epidemic diarrhea
Mixed infection, causes high mortality.
The method that preventing and treating pig wins card sick at present is more, mainly uses Western medicine, immunoprophylaxis, Chinese herbal medicine etc. to prevent and treat.Its
In, spreading unchecked that Western medicine uses brings great potential safety hazard to Pork food, and drug residue has become as affects Carnis Sus domestica product
One of principal element of product safety.Immunoprophylaxis early stage results is preferable, however it is necessary that and repeatedly inoculates, and troublesome poeration, cost is relatively
Height, easily causes stress；And vaccine requires higher for storage conditions, need cold preservation under the conditions of 2-8 DEG C, room
Under the conditions of temperature, a period of time then loses effect.Chinese herbal feed additive is also one of current research direction, but due in
The kind of medical herbs is more, and identical Chinese crude drug is very big to the difference between the effects of different diseases of bird and livestock, and disease is exempted from by Chinese herbal medicine
Epidemic disease effect is extremely limited, therefore applies and also has certain limitation.
Summary of the invention
It is an object of the invention to provide a kind of anti-pig and win the freeze-dried mixed powder that card is sick, solve prior art preventing and treating pig rich card disease and exist
Operation inconvenience, medicament be difficult to preserve, the poor problem of effect.
Second object of the present invention is to provide a kind of anti-pig and wins the preparation method of the sick freeze-dried mixed powder of card.
In order to realize object above, the technical solution adopted in the present invention is: a kind of anti-pig wins the freeze-dried mixed powder that card is sick, bag
Include the component of following parts by weight: the sick yolk antibody 94 of the rich card of anti-pig～97 parts, the sick transfer factor 1 of the rich card of anti-pig～2 parts,
Preservative 0.01～0.02 part, inactivator 0.2～0.3 part, selenium 0.05～0.1 part, vitamin C 0.5～1 part, polyethylene
Ketopyrrolidine 1～2 parts, glycine 0.5～1 part.
Described inactivator is formaldehyde.
Described preservative is thimerosal.
A kind of above-mentioned anti-pig wins the preparation method of the sick freeze-dried mixed powder of card, comprises the following steps:
A. using pig bocavirus type strain to prepare vaccine, immune health chicken group obtains high-immunity egg, high-immunity egg process is sterilized,
Acidification, centrifugal treating obtain winning the supernatant of the sick yolk antibody of card containing anti-pig；
B. by the supernatant sick transfer factor of card rich with anti-pig winning the sick yolk antibody of card containing anti-pig, preservative, inactivator, selenium,
Vitamin C, polyvinylpyrrolidone, glycine mix, then lyophilizing and get final product.
Preferably, anti-pig wins the preparation method of the sick freeze-dried mixed powder of card, comprises the following steps:
1) take pig bocavirus type strain to breed, inactivated and be prepared as vaccine, at interval of 7 days to healthy laying hen
Group carries out immunity, and immunity three times, start after immunity to detect antibody titer, when antibody titer reaches 1:256 for the second time altogether
Time, collect high-immunity egg, carry out the high-immunity egg of collection clearing up, sterilizing, dry standby；
2) by step 1) gained high-immunity egg isolated high-immunity egg yolk, add isopyknic sterilized water, be sufficiently stirred for,
To yolk antibody solution, yolk antibody solution adds acidified aqueous solution, mixes, staticly settle, take supernatant standby；
3) in step 2) gained supernatant adds sodium salt make sodium salt volume of the total volume 0.8%, centrifugation, must contain
Anti-pig is had to win the supernatant of the sick yolk antibody of card；
4) step 3 is taken) the anti-pig containing 94～97 weight portions of gained wins the supernatant of the sick yolk antibody of card, adds 1～2
The sick transfer factor of the rich card of the anti-pig of weight portion, filters, and reserved filtrate is standby；
5) by step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, adds preservative 0.01～0.02 weight portion, inactivation
Agent 0.2～0.3 weight portion, selenium 0.05～0.1 weight portion, vitamin C 0.5～1 weight portion, polyvinylpyrrolidone 1～2
Weight portion, glycine 0.5～1 weight portion mix, first-20 DEG C of pre-coolings, then lyophilizing under the conditions of-80 DEG C, to obtain final product.
Step 2) described in the pH of acidified aqueous solution be 4.9～5.2；The addition of acidified aqueous solution is yolk antibody solution body
Long-pending 8 times.Described acidified aqueous solution can be acidified aqueous solution.
Step 3) described in sodium salt be NaCl and Na2SO4The mixture that mass ratio is 4:1.
Step 3) described in the rotating speed of centrifugation be 15000rpm, the time is 20min.
Step 4) described in filter be first to filter with the filter membrane of 0.45 μm, molecular cut off dividing greatly more than 200kD
Son, then by filtrate with the membrane filtration of 0.22 μm.
The sick transfer factor of the rich card of described anti-pig, is prepared by following methods:
1), after pig spleen being carried out immunity with pig rich card sick virus antigen, its surface muscle, fat is removed, then with the life after sterilizing
Reason normal saline washing；
2), after the pig spleen after rinsing crushes, add the PBS after the pre-cooling of 2 times of volumes, use tissue homogenate instrument
It is homogeneous, obtain homogenate；
3), after homogenate being processed 5～10min with Ultrasonic Cell Disruptor, regulation pH is 5.5, at 5000～6000rpm rotating speeds
Under be centrifuged 30min, collect supernatant；
4) by step 3) gained supernatant micro-filtrate membrane filtration, molecular cut off is the material of more than 7000 dalton, will
Gained filtrate uses degerming membrane filtration, obtains the anti-pig sick transfer factor stock solution of rich card, saves backup under the conditions of 4 DEG C.
Yolk antibody is a kind of immunoglobulin, the one produced by the stimulation of external antigen due to body stress product,
Can combine with corresponding antigens, eliminate the cause of disease.Pig is won the chicken group that the sick vaccine virus immunization of card is healthy by the present invention, can make
The egg yolk obtained produces the effective anti-pig sick yolk antibody of rich card, decreases the probability of strain reversion simultaneously.
Transfer factor carries the specific immunity information of primed lymphocyte, it is possible to specific immunity information is presented to receptor
Lymphocyte, the lymphocyte making receptor inactive is changed into specificity primed lymphocyte, thus excites recipient cell to mediate
Immunoreation.
The anti-pig of the present invention is won in the freeze-dried mixed powder that card is sick, and the sick yolk antibody of the rich card of anti-pig is a kind of immunoglobulin, to pig
Rich card disease has good prevention effect；The sick transfer factor of the rich card of anti-pig itself is a kind of special antiviral agent, both is combined
Use, prevention can be greatly enhanced and treatment pig wins the effect that card is sick.
The anti-pig of the present invention wins the freeze-dried mixed powder that card is sick, the sick transfer factor of card rich with anti-pig of rich for the anti-pig sick yolk antibody of card is entered
Row is compounding, and is aided with preservative, inactivator, selenium, vitamin C, polyvinylpyrrolidone and glycine and prepares freeze-dried mixed
Powder, each component cooperates, coordinative role, can significantly improve its preventing and treating pig win card disease effect, have with strong points,
The advantage that room temperature lower holding time length, antibody titer are high, biological activity is good, is a kind of to have the potent of wide application prospect
Biological preparation；Proving through animal experiment, the anti-pig of the present invention wins the freeze-dried mixed powder that card is sick, and pig is won the prevention effect that card is sick
It is substantially better than that anti-pig wins the sick yolk antibody of card, anti-pig wins any one situation about being used alone in card disease transfer factor.
The anti-pig of the present invention wins the freeze-dried mixed powder that card is sick, during use, can press mass fraction 2%～3% as feed additive
It is mixed in feedstuff；Intramuscular injection mode is used to be administered during morbidity, containing freeze-dried mixed powder 2g in every 100ml injection, injection
Usage amount is each 0.1～0.2ml/kg body weight；Medication first doubles, and therapeutic effect is more preferable.
The anti-pig of the present invention wins the preparation method of the sick freeze-dried mixed powder of card, by rich for the anti-pig sick yolk antibody of card, anti-pig rich card disease
Transfer factor and multiple auxiliary materials compound, and by adjusting the composition of adjuvant, controlling its consumption, extend yolk further
The holding time of antibody, it is ensured that the activity in the yolk antibody long period at normal temperatures, reduce the preservation of freeze-dried powder preparation
And cost of transportation, easy to use, it is with a wide range of applications；Preparation method technique is simple, easy to operate, drops greatly
Low production cost, is suitable for large-scale industrial production.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention is further illustrated.
In the detailed description of the invention of the present invention, pig bocavirus used is national standard strain.
In the detailed description of the invention of the present invention, the sick transfer factor of the rich card of anti-pig used, following methods prepare:
1), after pig spleen being carried out immunity with pig rich card sick Virus Standard antigen (commercially available), its surface muscle, fat is removed, then
With the normal saline flushing after sterilizing；
2), after the pig spleen after rinsing crushes, add the PBS after the pre-cooling of 2 times of volumes, use tissue homogenate instrument
It is homogeneous, obtain homogenate；
3), after homogenate being processed 5～10min with Ultrasonic Cell Disruptor, regulation pH is 5.5, at 5000～6000rpm rotating speeds
Under be centrifuged 30min, collect supernatant；
4) by step 3) gained supernatant micro-filtrate membrane filtration, molecular cut off is the material of more than 7000 dalton, will
Gained filtrate uses degerming membrane filtration, obtains the anti-pig sick transfer factor stock solution of rich card, saves backup under the conditions of 4 DEG C.
Embodiment 1
The anti-pig of the present embodiment wins the freeze-dried mixed powder that card is sick, including the component of following parts by weight: the sick yolk of the rich card of anti-pig resists
Body 94 parts, the rich card of anti-pig sick transfer factor 2 parts, thimerosal 0.02 part, 0.2 part of formaldehyde, 0.05 part of selenium, vitamin C 1
Part, polyvinylpyrrolidone 2 parts, glycine 0.5 part.
The anti-pig of the present embodiment wins the preparation method of the sick freeze-dried mixed powder of card, comprises the following steps:
1) take pig bocavirus standard strain to breed, inactivated and be prepared as vaccine, at interval of 7 days, health is laid eggs
Chicken group carries out immunity, altogether immunity three times, and immune health chicken group inoculates inactivated vaccine 1.5ml for the first time, for the second time, the 3rd
Secondary immunity inoculation dosage 1 times, starts after immunity to detect antibody titer for the second time, when antibody titer reaches 1:256, receives
Collection high-immunity egg, carries out the high-immunity egg of collection clearing up, sterilizing, dries standby；
2) by step 1) gained high-immunity egg isolated high-immunity egg yolk, add isopyknic sterilized water, be sufficiently stirred for,
To yolk antibody solution, adding pH in yolk antibody solution is the acidified aqueous solution of 5.0, and acidified aqueous solution addition is ovum
8 times of yellow antibody-solutions volume, mix, staticly settle, take supernatant standby；
3) in step 2) gained supernatant adds by NaCl and Na2SO4Ratio for 4:1 in mass ratio is mixed to form
Mixing sodium salt, makes mixing sodium salt volume of the total volume 0.8%, is centrifuged 20min with the rotating speed of 15000rpm, must be containing anti-
Pig wins the supernatant of the sick yolk antibody of card；
4) step 3 is taken) the anti-pig containing 94 weight portions of gained wins the supernatant of the sick yolk antibody of card, adds 2 weight portions
The sick transfer factor of the rich card of anti-pig, filters with the filter membrane of 0.45 μm, the molecular weight that the dams macromolecular substances more than 200kD,
Again by filtrate through 0.22 μm membrane filtration, reserved filtrate is standby；
5) by step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, adds thimerosal 0.02 weight portion, formaldehyde 0.2 weight
Amount part, selenium 0.05 weight portion, vitamin C 1 weight portion, polyvinylpyrrolidone 2 weight portion, glycine 0.5 weight portion
Mix, first-20 DEG C of pre-coolings, then lyophilizing under the conditions of-80 DEG C, to obtain final product.
Embodiment 2
The anti-pig of the present embodiment wins the freeze-dried mixed powder that card is sick, including the component of following parts by weight: the sick yolk of the rich card of anti-pig resists
Body 95 parts, the rich card of anti-pig sick transfer factor 1.5 parts, thimerosal 0.01 part, 0.3 part of formaldehyde, 0.1 part of selenium, vitamin C 0.5
Part, polyvinylpyrrolidone 1.5 parts, glycine 1 part.
The anti-pig of the present embodiment wins the preparation method of the sick freeze-dried mixed powder of card, comprises the following steps:
1) take pig bocavirus standard strain to breed, inactivated and be prepared as vaccine, at interval of 7 days, health is laid eggs
Chicken group carries out immunity, altogether immunity three times, and immune health chicken group inoculates inactivated vaccine 1.5ml for the first time, for the second time, the 3rd
Secondary immunity inoculation dosage 1 times, starts after immunity to detect antibody titer for the second time, when antibody titer reaches 1:256, receives
Collection high-immunity egg, carries out the high-immunity egg of collection clearing up, sterilizing, dries standby；
2) by step 1) gained high-immunity egg isolated high-immunity egg yolk, add isopyknic sterilized water, be sufficiently stirred for,
To yolk antibody solution, adding pH in yolk antibody solution is the acidified aqueous solution of 5.0, and acidified aqueous solution addition is ovum
8 times of yellow antibody-solutions volume, mix, staticly settle, take supernatant standby；
3) in step 2) gained supernatant adds by NaCl and Na2SO4Ratio for 4:1 in mass ratio is mixed to form
Mixing sodium salt, makes mixing sodium salt volume of the total volume 0.8%, is centrifuged 20min with the rotating speed of 15000rpm, must be containing anti-
Pig wins the supernatant of the sick yolk antibody of card；
4) step 3 is taken) the anti-pig containing 95 weight portions of gained wins the supernatant of the sick yolk antibody of card, adds 1.5 weight
The sick transfer factor of the rich card of the anti-pig of part, filters with the filter membrane of 0.45 μm, the molecular weight that the dams macromolecular complex more than 200kD
Matter, then by filtrate through 0.22 μm membrane filtration, reserved filtrate is standby；
5) by step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, adds thimerosal 0.01 weight portion, formaldehyde 0.3 weight
Amount part, selenium 0.1 weight portion, vitamin C 0.5 weight portion, polyvinylpyrrolidone 1.5 weight portion, glycine 1 weight portion
Mix, first-20 DEG C of pre-coolings, then lyophilizing under the conditions of-80 DEG C, to obtain final product.
Embodiment 3
The anti-pig of the present embodiment wins the freeze-dried mixed powder that card is sick, including the component of following parts by weight: the sick yolk of the rich card of anti-pig resists
Body 97 parts, the rich card of anti-pig sick transfer factor 1 part, thimerosal 0.01 part, 0.25 part of formaldehyde, 0.08 part of selenium, vitamin C
1 part, polyvinylpyrrolidone 1 part, glycine 0.5 part.
The anti-pig of the present embodiment wins the preparation method of the sick freeze-dried mixed powder of card, comprises the following steps:
1) take pig bocavirus standard strain to breed, inactivated and be prepared as vaccine, at interval of 7 days, health is laid eggs
Chicken group carries out immunity, altogether immunity three times, and immune health chicken group inoculates inactivated vaccine 1.5ml for the first time, for the second time, the 3rd
Secondary immunity inoculation dosage 1 times, starts after immunity to detect antibody titer for the second time, when antibody titer reaches 1:256, receives
Collection high-immunity egg, carries out the high-immunity egg of collection clearing up, sterilizing, dries standby；
2) by step 1) gained high-immunity egg isolated high-immunity egg yolk, add isopyknic sterilized water, be sufficiently stirred for,
To yolk antibody solution, adding pH in yolk antibody solution is the acidified aqueous solution of 5.0, and acidified aqueous solution addition is ovum
8 times of yellow antibody-solutions volume, mix, staticly settle, take supernatant standby；
3) in step 2) gained supernatant adds by NaCl and Na2SO4Ratio for 4:1 in mass ratio is mixed to form
Mixing sodium salt, makes mixing sodium salt volume of the total volume 0.8%, is centrifuged 20min with the rotating speed of 15000rpm, must be containing anti-
Pig wins the supernatant of the sick yolk antibody of card；
4) step 3 is taken) the anti-pig containing 97 weight portions of gained wins the supernatant of the sick yolk antibody of card, adds 1 weight portion
The sick transfer factor of the rich card of anti-pig, filters with the filter membrane of 0.45 μm, the molecular weight that the dams macromolecular substances more than 200kD,
Again by filtrate through 0.22 μm membrane filtration, reserved filtrate is standby；
5) by step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, adds thimerosal 0.01 weight portion, formaldehyde 0.25 weight
Amount part, selenium 0.08 weight portion, vitamin C 1 weight portion, polyvinylpyrrolidone 1 weight portion, glycine 0.5 weight portion
Mix, first-20 DEG C of pre-coolings, then lyophilizing under the conditions of-80 DEG C, to obtain final product.
Experimental example 1
This experimental example carries out performance detection to the embodiment 1～3 freeze-dried mixed powder of gained, and result is as shown in table 1,2.
Room temperature bioactivity experimental technique: the embodiment 1～3 freeze-dried mixed powder preparation of gained is saved in 37 DEG C of incubators,
Preserve 24 months, respectively 1 week, 2 weeks, January, February, April, August, December, 16 months, 20 months, after 24 months,
By agar diffusion (AGP) method, measure the yolk antibody titer change of the taken sample of each time point.
Table 1 embodiment 1～3 gained freeze-dried mixed powder room temperature bioactivity experimental result (project: PRRS)
| Group | 1 week | 2 weeks | January | February | April | August | December | 16 months | 20 months | 24 months |
| Embodiment 1 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1；16 | 1:16 | 1:16 | 1:16 |
| Embodiment 2 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1；16 | 1:16 | 1:16 | 1:16 |
| Embodiment 3 | 1:32 | 1:32 | 1:32 | 1:32 | 1:32 | 1:16 | 1；16 | 1:16 | 1:16 | 1:16 |
Table 2 embodiment 1～3 gained freeze-dried mixed powder room temperature places performance test results
Experimental example 2
This experimental example the anti-pig of embodiment 1～3 is won the sick freeze-dried mixed powder preventing and treating pig of card win the sick effect of card carry out clinical in fact
Test.
Experimental technique: choosing 15 age in days pig 700, wherein 100 be only used as counteracting toxic substances group isolated rearing, 100 are only used as blank
Group isolated rearing, 200 are only used as matched group is divided into 2 groups, often group 100, isolated rearing, respectively by comparative example 1,2
Preparation for treating, other 300 are only used as test group is divided into 3 groups, often group 100, respectively with the preparation of embodiment 1,2,3
Treatment.Feeding manner is free choice feeding normal diet, freely drinks water, and after 24h, takes test group, matched group and counteracting toxic substances group
Pig bocavirus liquid after pig intramuscular injection inoculation dilution, inoculum concentration is 2ml/；Beginning for property treatment after infection, intramuscular injection,
Injection dosage is 0.2ml/kg body weight；Death condition after record 120h, result is as shown in table 3.
Pig is won the prevention effect experimental result that card is sick by table 3 the embodiment 1～3 freeze-dried mixed powder of gained
The medicament of comparative example 1 is made up of the component of following parts by weight: the rich card of anti-pig sick yolk antibody 97 parts, thimerosal 0.01
Part, 0.25 part of formaldehyde, 0.08 part of selenium, vitamin C 0.8 part, polyvinylpyrrolidone 1.5 parts, glycine 0.5 part.
The medicament of comparative example 2 is made up of the component of following parts by weight: the rich card of anti-pig sick transfer factor 2 parts, thimerosal 0.01
Part, 0.25 part of formaldehyde, 0.08 part of selenium, vitamin C 0.8 part, polyvinylpyrrolidone 1.5 parts, glycine 0.5 part.
From table 3 it is observed that the embodiment 1～3 freeze-dried mixed powder of gained can effectively prevent and treat pig rich card disease, and mixing is frozen
Dry powder effect to be substantially better than that anti-pig wins the sick yolk antibody of card, anti-pig wins any one feelings being used alone in card disease transfer factor
Condition.
Claims (9)
1. an anti-pig wins the freeze-dried mixed powder that card is sick, it is characterised in that: include the component of following parts by weight: anti-pig is won
The sick yolk antibody 94 of card～97 parts, the sick transfer factor 1 of rich card of anti-pig～2 parts, preservative 0.01～0.02 part, inactivator 0.2～
0.3 part, selenium 0.05～0.1 part, vitamin C 0.5～1 part, polyvinylpyrrolidone 1～2 parts, glycine 0.5～1 part；
Described freeze-dried mixed powder is to be prepared by the method comprised the following steps:
A. using pig bocavirus type strain to prepare vaccine, immune health chicken group obtains high-immunity egg, high-immunity egg process is sterilized,
Acidification, centrifugal treating obtain winning the supernatant of the sick yolk antibody of card containing anti-pig；
B. by the supernatant sick transfer factor of card rich with anti-pig winning the sick yolk antibody of card containing anti-pig, preservative, inactivator, selenium,
Vitamin C, polyvinylpyrrolidone, glycine mix, then lyophilizing and get final product；
Wherein, the sick transfer factor of the rich card of described anti-pig, following methods prepare:
1), after pig spleen being carried out immunity with pig rich card sick virus antigen, its surface muscle, fat is removed, then with the life after sterilizing
Reason normal saline washing；
2), after the pig spleen after rinsing crushes, add the PBS after the pre-cooling of 2 times of volumes, use tissue homogenate instrument
It is homogeneous, obtain homogenate；
3), after homogenate being processed 5～10min with Ultrasonic Cell Disruptor, regulation pH is 5.5, at 5000～6000rpm rotating speeds
Under be centrifuged 30min, collect supernatant；
4) by step 3) gained supernatant micro-filtrate membrane filtration, molecular cut off is the material of more than 7000 dalton, will
Gained filtrate uses degerming membrane filtration, obtains the anti-pig sick transfer factor stock solution of rich card, saves backup under the conditions of 4 DEG C.
Anti-pig the most according to claim 1 wins the freeze-dried mixed powder that card is sick, it is characterised in that: described inactivator is first
Aldehyde.
Anti-pig the most according to claim 1 wins the freeze-dried mixed powder that card is sick, it is characterised in that: described preservative is sulfur
Willow hydrargyrum.
4. an anti-pig as claimed in claim 1 wins the preparation method of the sick freeze-dried mixed powder of card, it is characterised in that: bag
Include the following step:
A. using pig bocavirus type strain to prepare vaccine, immune health chicken group obtains high-immunity egg, high-immunity egg process is sterilized,
Acidification, centrifugal treating obtain winning the supernatant of the sick yolk antibody of card containing anti-pig；
B. by the supernatant sick transfer factor of card rich with anti-pig winning the sick yolk antibody of card containing anti-pig, preservative, inactivator, selenium,
Vitamin C, polyvinylpyrrolidone, glycine mix, then lyophilizing and get final product.
Anti-pig the most according to claim 4 wins the preparation method of the sick freeze-dried mixed powder of card, it is characterised in that: include
The following step:
1) take pig bocavirus type strain to breed, inactivated and be prepared as vaccine, at interval of 7 days to healthy laying hen
Group carries out immunity, and immunity three times, start after immunity to detect antibody titer, when antibody titer reaches 1:256 for the second time altogether
Time, collect high-immunity egg, carry out the high-immunity egg of collection clearing up, sterilizing, dry standby；
2) by step 1) gained high-immunity egg isolated high-immunity egg yolk, add isopyknic sterilized water, be sufficiently stirred for,
To yolk antibody solution, yolk antibody solution adds acidified aqueous solution, mixes, staticly settle, take supernatant standby；
3) in step 2) gained supernatant adds sodium salt make sodium salt volume of the total volume 0.8%, centrifugation, must contain
Anti-pig is had to win the supernatant of the sick yolk antibody of card；
4) step 3 is taken) the anti-pig containing 94～97 weight portions of gained wins the supernatant of the sick yolk antibody of card, adds 1～2
The sick transfer factor of the rich card of the anti-pig of weight portion, filters, and reserved filtrate is standby；
5) by step 4) gained filtrate is concentrated into 2.5ml/500 plumage part, adds preservative 0.01～0.02 weight portion, inactivation
Agent 0.2～0.3 weight portion, selenium 0.05～0.1 weight portion, vitamin C 0.5～1 weight portion, polyvinylpyrrolidone 1～2
Weight portion, glycine 0.5～1 weight portion mix, first-20 DEG C of pre-coolings, then lyophilizing under the conditions of-80 DEG C, to obtain final product.
Anti-pig the most according to claim 5 wins the preparation method of the sick freeze-dried mixed powder of card, it is characterised in that: step 2)
Described in the pH of acidified aqueous solution be 4.9～5.2；The addition of acidified aqueous solution is 8 times of yolk antibody liquor capacity.
Anti-pig the most according to claim 5 wins the preparation method of the sick freeze-dried mixed powder of card, it is characterised in that: step 3)
Described in sodium salt be NaCl and Na2SO4The mixture that mass ratio is 4:1.
Anti-pig the most according to claim 5 wins the preparation method of the sick freeze-dried mixed powder of card, it is characterised in that: step 3)
Described in the rotating speed of centrifugation be 15000rpm, the time is 20min.
Anti-pig the most according to claim 5 wins the preparation method of the sick freeze-dried mixed powder of card, it is characterised in that: step 4)
Described in filter be first to filter with the filter membrane of 0.45 μm, molecular cut off is more than the macromole of 200kD, then by filtrate
Membrane filtration by 0.22 μm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310463389.9A CN104208680B (en) | 2013-09-30 | 2013-09-30 | A kind of anti-pig wins sick freeze-dried mixed powder of card and preparation method thereof |
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| CN201310463389.9A CN104208680B (en) | 2013-09-30 | 2013-09-30 | A kind of anti-pig wins sick freeze-dried mixed powder of card and preparation method thereof |
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| Publication Number | Publication Date |
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| CN104208680A CN104208680A (en) | 2014-12-17 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101096704A (en) * | 2006-06-01 | 2008-01-02 | 杭州致远医学检验所有限公司 | A set of fluorescent quantitative PCR primer and probe for detecting human boca virus |
| CN102908620A (en) * | 2011-08-04 | 2013-02-06 | 广州格拉姆生物科技有限公司 | Preparation method of egg yolk antibody injection for treating piglet diarrhea |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101096704A (en) * | 2006-06-01 | 2008-01-02 | 杭州致远医学检验所有限公司 | A set of fluorescent quantitative PCR primer and probe for detecting human boca virus |
| CN102908620A (en) * | 2011-08-04 | 2013-02-06 | 广州格拉姆生物科技有限公司 | Preparation method of egg yolk antibody injection for treating piglet diarrhea |
Non-Patent Citations (2)
| Title |
|---|
| 人博卡病毒结构和非结构蛋白的原核表达、纯化及多克隆抗体的制备;王雅英等;《中国生物制品学杂质》;20110531;第24卷(第5期);第497-501页 * |
| 人类博卡病毒(HBoV)非结构蛋白NS1基因的克隆、原核表达及多克隆抗体的制备;孙彬等;《现代生物医学进展》;20100331;第10卷(第6期);第1013-1016页 * |
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