CN101376026A - Method for preparing vaccine of porcine circovirus type 2 - Google Patents
Method for preparing vaccine of porcine circovirus type 2 Download PDFInfo
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- CN101376026A CN101376026A CNA2008100795605A CN200810079560A CN101376026A CN 101376026 A CN101376026 A CN 101376026A CN A2008100795605 A CNA2008100795605 A CN A2008100795605A CN 200810079560 A CN200810079560 A CN 200810079560A CN 101376026 A CN101376026 A CN 101376026A
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Abstract
The invention discloses a method for preparing a porcine circovirus type 2 virus vaccine. Viscera tissues are separated out from a sick pig body with typical circovirus type 2 virus, and an extractive of viscera tissues is obtained after rubbing and filtration of the viscera tissues; after a formaldehyde inactivation is added, the extractive is diluted by physiological saline and prepared into an original inactivation vaccine; after a healthy pig body is inoculated with the inactivation vaccine and is led to its onset or death, the viscera tissues are collected, and after the rubbing and filtration, and with the added formaldehyde inactivation, the viscera tissues are diluted by physiological saline and prepared into the porcine circovirus type 2 virus vaccine. The porcine circovirus type 2 virus vaccine prepared by the method has the advantages of a good antigenicity and a high immunity protection rate, thus solving the problems of the extremely difficult virus multiplication and high production cost in the prior production of the porcine circovirus type 2 virus vaccine.
Description
Technical field
The invention belongs to the biovaccine preparing technical field, relate to the preparation method of a boar, particularly relate to the preparation method of a boar 2 type porcine circovirus vaccines with vaccine.
Background technology
Pig 2 type porcine circovirus (PCV2) are a kind of new pig infectious disease, mainly cause ablactational baby pig multisystemic wasting syndrome and nephropathy dermatitis syndrome, also can produce immunosuppressant to a certain extent simultaneously, cause the secondary and the accompanying infection of other disease.The nineties in 20th century, Canada broke out this disease first, and all there is this pathogenetic report many in the world subsequently countries and regions, have caused serious economy loss to pig industry.PCV2 is a kind of DNA viruses, the aberration rate of virus is low, in cell culture, do not produce cytometaplasia, be difficult to produce the virion of high titre, it is very inconvenient to prepare the PCV2 vaccine with routine techniques, therefore, does not all have the conventional vaccine listing of pig 2 type porcine circovirus so far both at home and abroad, shortage does not have effective medicine and effective control ability to this disease simultaneously to effective prophylactico-therapeutic measures of this disease yet.Along with development of science and technology, people constantly increase for the pig organ being used for the heteroplastic interest of philtrum, and present present situation has caused potential public health risks to the people virtually.
Chinese patent CN101240264A " porcine circovirus 2 type inactivated vaccine " discloses a kind of from sick pig farm collection pathological material of disease, after carrying out virus separation, purification, be seeded in the PK-15 cell and breed in a large number, then through formalin-inactivated, add white-oil adjuvant emulsifying, be prepared into the method for conventional white-oil adjuvant vaccine.To be that the propagation difficulty of virus is big, cost is high, be difficult to supply in a large number vaccine satisfy the production demand to the problem that this technology exists.
Summary of the invention
The preparation method that the purpose of this invention is to provide a boar 2 type porcine circovirus vaccines, the responsive internal organs that utilize the disease pig are as the viral source material, make pig 2 type porcine circovirus inactivated vaccines by inactivation technology, to solve the problem that to prevent pig 2 type porcine circovirus at present.
The preparation method of pig 2 type porcine circovirus vaccines of the present invention is:
1). 2 type porcine circovirus pigs of the typical case 2 type porcine circovirus pigs that selection has just been died of illness or typical case's morbidity, under aseptic condition, in its body, isolate organs and tissues, grind with colloid mill, remove by filter impurity, obtain the organs and tissues extract, according to the organs and tissues extract: the volume ratio of formaldehyde=40: 1 adds formaldehyde, deactivation is 5 days under 30 ℃ of conditions, filtration sterilization, the volumetric concentration that is diluted to the organs and tissues extract with normal saline is 40%, make original inactivated vaccine, deposit under 2～8 ℃ of environment standby;
2). above-mentioned vaccination in the health pig body, is impelled its morbidity;
3). when the health pig of inoculation is typical 2 type porcine circovirus morbidity or death, under aseptic condition, gather organs and tissues, grind with colloid mill, remove by filter impurity, obtain the organs and tissues extract, according to the organs and tissues extract: the volume ratio of formaldehyde=40: 1 adds formaldehyde, deactivation is 5 days under 30 ℃ of conditions, filtration sterilization, the volumetric concentration that is diluted to the organs and tissues extract with normal saline is 40%, make pig 2 type porcine circovirus inactivated vaccines, be divided in the sterilization bottle, deposit under 2～8 ℃ of environment standby.
Above-mentioned organs and tissues extract is meant under aseptic condition, the extract of the internal organs heart, liver, spleen, lung and the kidney of gathering in typical pig 2 type porcine circovirus morbidity or the dead pig body.
The percent by volume of the pig 2 type porcine circovirus vaccines that the present invention prepares consists of:
Formaldehyde 1%
Organs and tissues extract 40%
Normal saline 59%.
If make original vaccine, still be inoculated in this pig farm prevention pig 2 type porcine circovirus, then better effects if behind the production pig 2 type porcine circovirus inactivated vaccines with the pig that dies of illness in the pig farm.
The using method of pig 2 type porcine circovirus vaccines of the present invention is: when piglet 5 ages in days, and every pig muscle injection pig 2 type porcine circovirus inactivated vaccine 2mL.
Select 20 conducts of 5 healthy age in days piglets for the examination object, be divided into two groups, 10 every group.Every pig muscle injection pig of immune group 2 type porcine circovirus inactivated vaccine 2mL, matched group is immunoprophylaxis not, observes its health status.
Gather the die of illness internal organs of pig of typical 2 type porcine circovirus, after grinding with colloid mill, make pig 2 type porcine circovirus extracting solution.
After 25 days, to test piggy injection pig 2 type porcine circovirus extracting solution, two groups of pigs are all injected, and every pig muscle injection 1mL carries out strong virus attack, observes the disease of pig, dead situation.
Result of the test shows that in whole 10 piglets of immune group, 8 strong living do not have disease symptom, and 2 have only disease symptom, and no death condition takes place; 10 piglets of matched group all fall ill, dead 6.Immune group is compared with matched group, and immune protective rate reaches 80%.
The present invention utilize the disease pig each responsive internal organs as original inactivated vaccine; inoculate to the health pig body and produce preparation pig 2 type porcine circovirus inactivated vaccines; the vaccine antigen of preparing is good; the immune protective rate height has solved the problem that the virus multiplication difficulty is big, production cost is high in the present pig 2 type porcine circovirus production of vaccine.
Pig 2 type porcine circovirus inactivated vaccine preparation methoies of the present invention are simple, easy to use, easy to utilize.
The specific embodiment
Embodiment 1
Under the aseptic condition, in the typical pig 2 type circovirus disease pig bodies of just having died of illness, take out organs and tissues such as the heart, liver, spleen, lung and kidney, fully grind, remove by filter impurity, obtain the organs and tissues extract with colloid mill.Taking internal organ is organized 4mL, adds 0.1mL formaldehyde, and deactivation is 5 days under 30 ℃ of conditions, through the qualified after-filtration degerming of inspection of going out, adds 5.9mL normal saline mix homogeneously, by steriling test, efficacy test and safety verification, makes original inactivated vaccine.
In original inactivated vaccine intramuscular injection to a health pig body, impel its morbidity, when the health pig of inoculation is the morbidity of typical pig 2 type circovirus disease, under aseptic condition, gather organs and tissues such as its heart, liver, spleen, lung and kidney, fully grind with colloid mill, remove by filter impurity, obtain the organs and tissues extract.Taking internal organ is organized 800mL, add 20mL formaldehyde, deactivation is 5 days under 30 ℃ of conditions, go out and examine qualified after-filtration degerming, add 1180mL normal saline mix homogeneously,, be mixed with pig 2 type porcine circovirus inactivated vaccines by steriling test, efficacy test and safety verification, be divided in the 100mL bottle of sterilization, under 2～8 ℃ condition, deposit standby.
Embodiment 2
Under the aseptic condition, in the typical pig 2 type circovirus disease pig bodies of typical case's morbidity, take out organs and tissues such as the heart, liver, spleen, lung and kidney, fully grind, remove by filter impurity, obtain the organs and tissues extract with colloid mill.Taking internal organ is organized 4mL, adds 0.1mL formaldehyde, and deactivation is 5 days under 30 ℃ of conditions, through the qualified after-filtration degerming of inspection of going out, adds 5.9mL normal saline mix homogeneously, by steriling test, efficacy test and safety verification, makes original inactivated vaccine.
In original inactivated vaccine intramuscular injection to a health pig body, impel its morbidity, after the health pig of inoculation is typical pig 2 type circovirus disease morbidity death, under aseptic condition, gather organs and tissues such as its heart, liver, spleen, lung and kidney, grind with colloid mill, remove by filter impurity, obtain the organs and tissues extract.Taking internal organ is organized 800mL, add 20mL formaldehyde, deactivation is 5 days under 30 ℃ of conditions, go out and examine qualified after-filtration degerming, add 1180mL normal saline mix homogeneously,, be mixed with pig 2 type porcine circovirus inactivated vaccines by steriling test, efficacy test and safety verification, be divided in the 100mL bottle of sterilization, under 2～8 ℃ condition, deposit standby.
Claims (3)
1, the preparation method of a boar 2 type porcine circovirus vaccines is with following step preparation:
1). 2 type porcine circovirus pigs of the typical case 2 type porcine circovirus pigs that selection has just been died of illness or typical case's morbidity, under aseptic condition, in its body, isolate organs and tissues, grind with colloid mill, remove by filter impurity, obtain the organs and tissues extract, according to the organs and tissues extract: the volume ratio of formaldehyde=40: 1 adds formaldehyde, deactivation is 5 days under 30 ℃ of conditions, filtration sterilization, the volumetric concentration that is diluted to the organs and tissues extract with normal saline is 40%, make original inactivated vaccine, deposit under 2～8 ℃ of environment standby;
2). above-mentioned vaccination in the health pig body, is impelled its morbidity;
3). when the health pig of inoculation is typical 2 type porcine circovirus morbidity or death, under aseptic condition, gather organs and tissues, grind with colloid mill, remove by filter impurity, obtain the organs and tissues extract, according to the organs and tissues extract: the volume ratio of formaldehyde=40: 1 adds formaldehyde, deactivation is 5 days under 30 ℃ of conditions, filtration sterilization, the volumetric concentration that is diluted to the organs and tissues extract with normal saline is 40%, make pig 2 type porcine circovirus inactivated vaccines, be divided in the sterilization bottle, deposit under 2～8 ℃ of environment standby.
2, the preparation method of pig 2 type porcine circovirus vaccines according to claim 1, it is characterized in that described organs and tissues extract is meant under aseptic condition, the extract of the internal organs heart, liver, spleen, lung and the kidney of gathering in morbidity of typical pig 2 type porcine circovirus or the dead pig body.
3, the pig 2 type porcine circovirus vaccines that prepare of the preparation method of the described pig 2 type porcine circovirus vaccines of claim 1, its percent by volume consists of:
Formaldehyde 1%
Organs and tissues extract 40%
Normal saline 59%.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100795605A CN101376026A (en) | 2008-10-10 | 2008-10-10 | Method for preparing vaccine of porcine circovirus type 2 |
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| CNA2008100795605A CN101376026A (en) | 2008-10-10 | 2008-10-10 | Method for preparing vaccine of porcine circovirus type 2 |
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| CN101376026A true CN101376026A (en) | 2009-03-04 |
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| CNA2008100795605A Pending CN101376026A (en) | 2008-10-10 | 2008-10-10 | Method for preparing vaccine of porcine circovirus type 2 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102716476A (en) * | 2012-05-31 | 2012-10-10 | 郑州后羿制药有限公司 | Cell inactivated vaccine, egg yolk antibody injection, and preparation method of cell inactivated vaccine |
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- 2008-10-10 CN CNA2008100795605A patent/CN101376026A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102716476A (en) * | 2012-05-31 | 2012-10-10 | 郑州后羿制药有限公司 | Cell inactivated vaccine, egg yolk antibody injection, and preparation method of cell inactivated vaccine |
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Open date: 20090304 |