CN102735808A - Effect testing method for duck hemorrhagic ovaritis inactivated vaccine - Google Patents
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Abstract
The present invention provides an effect testing method for a duck hemorrhagic ovaritis inactivated vaccine. According to the method, a duck hemorrhagic ovaritis inactivated vaccine is adopted to vaccinate ducks twice, the second time vaccination is performed after the first time vaccination is performed two weeks, duck hemorrhagic ovaritis virus is adopted to carry out virus challenge on the vaccinated ducks after the second time vaccination is performed 3-5 weeks, the tested ducks are killed after the virus challenge is performed 5-10 days, autopsy is performed, and general pathological changes of a genital system are observed, wherein the pathological changes occur if the female ducks present ovarian follicle bleeding, deformation and oviduct atrophy, or the male ducks present testis atrophy and deferent duct atrophy, and the qualified standard of the vaccine effect test is that the virus challenge protection rate of the vaccinated ducks without pathological changes of the genital system is more than 70%.
Description
Technical field
The present invention relates to a kind of method for testing efficacy of inactivated vaccine for animals, relate in particular to the method for inspection that a kind of duck hemorrhagic oaritis inactivated vaccine is renderd a service, belong to veterinary biologics inspection technology field.
Background technology
Duck hemorrhagic oaritis (Duck Hemorrhagic Ovaritis; DH) be a kind of novel duck infectious disease popular recently, acute, the height contact, its cause of disease be duck hemorrhagic oaritis virus (Duck Hemorrhagic Ovaritis Virus, DHOV); The member who on virus taxis, belongs to flaviviridae; Infect the different days duck, cause that the ovarian hemorrhage of sexal maturity duck, orchiatrophy, egg production, rate of fertilization and incubation rate descend, the duck death rate increases.
In the research of virus taxis status; Through the F4 of isolated viral strain is carried out sequence homology analysis and evolutionary analysis for the go down to posterity genome sequence of virus and known virus of duck embryo, find that the genetic distance of tembusu virus of this virus and flaviviridae Flavivirus is nearest.Aspect diagnosis, people's report can adopt clinical symptoms such as Cao Zhenzhen, Wan Chun and, Su Jingliang, Teng Qiao are vast, reach histopathology variation, pcr amplification and virus substantially and method such as separate and make a definite diagnosis this disease.About these sick prevention and control, the strain that Beijing City Agriculture and Forestry Institute and Ruipu (Baoding) Biological Pharmaceutical Co., Ltd., Qilu Animal Health Products Co., Ltd., Fujian Province Academy Of Agricultural Sciences Animal Husbandry And Veterinary Medicine Institute and academy of agricultural sciences, Chongqing City animal and veterinary research institute have been separated during having reported and having utilized this disease to break out prepares the method for vaccine.Lin Jian, Liu Yuehuan etc. have reported the artificial challenge's model that utilizes laying ducks successfully to set up duck hemorrhagic oaritis, for experiment basis has been established in the evaluation of vaccine.That Shang Weijian has is objective, feasible, the play-by-play of the vaccine potency evaluation method of strong operability and science.
Summary of the invention
One of the object of the invention provides age in days scope and the sex of a kind of duck hemorrhagic oaritis inactivated vaccine effect evaluation duck.
Two of the object of the invention provide duck hemorrhagic oaritis inactivated vaccine effect evaluation with the tissue samples of vaccine immunity number of times, employing time, collection, attack malicious approach and embryo is used in check.
Three of the object of the invention provides a kind of duck hemorrhagic oaritis inactivated vaccine effect evaluation method.
The used virus stain called after of the present invention duck hemorrhagic oaritis virus HB strain, preserving number is CCTCC V201122; Said duck hemorrhagic oaritis virus is ssRNA virus, and cyst membrane is arranged, can aggegation dove red blood cell; It is spherical that virion is substantially, diameter 40～60nm; The said virus resistibility of environment to external world is stronger, in 4 ℃, deposits several weeks, in-20 ℃, deposits some months, and its appeal is all unaffected; Said virus is responsive to ether, chloroform; Most of detergents can be with its rapid deactivation; Under 37 ℃ of conditions, stifling 6h just can be its deactivation with 0.1% formalin.This virus can be at 9～13 age in days duck embryo allantoic cavities, embryo CAM, 9～10 age in days chick embryo allantoic cavities, embryo CAM, and 6 age in days chick embryo yolk sacs are cultivated in 7～8 age in days duck embryo yolk bags, also can on DEF and BHK21 cell, cultivate.
The preparation method of the used duck hemorrhagic of the present invention oaritis inactivated vaccine has declared patent of invention in 2011, application number is 201110255115.1.
The present invention provides a kind of duck hemorrhagic oaritis inactivated vaccine method for testing efficacy; This method is used duck hemorrhagic oaritis inactivated vaccine immune duck; Immunity is 2 times altogether, carries out 22 weeks after head exempts from and exempts from, and two exempt from 3 ~ 5 weeks of back attacks poison with duck hemorrhagic oaritis virus to immune duck; Attack the poison back and will test duck execution on the 5th～10, cut open inspection, observe the cardinal principle pathology of reproductive system, the malicious protection ratio of attacking of pathology does not take place the reproductive organs of immune group duck is to be judged to vaccine potency more than 70% to be up to the standards; Described cardinal principle pathology shows as duck that the duck ovarian follicle is hemorrhage, distortion or fallopian tubal atrophy, shows as testis or atrophy of vas deferens male duck.
Further, the inventive method confirms that according to the neurological susceptibility experimental result it is 120 ~ 560 ages in days that duck hemorrhagic oaritis inactivated vaccine efficacy test uses the age in days of duck.
Preferably, the sex of efficacy test use duck is female.
Wherein, the dosage of 2 immune duck hemorrhagic oaritis inactivated vaccines be 0.5ml/ only～2ml/ only.
Wherein, attack poison and uses duck hemorrhagic oaritis virus to be DHOV-HB strain (deposit number is CCTCC V201122, has been to disclose in 201110255115.1 the Chinese patent at application number), attacking the toxic agent amount is 3 * 10
4ELD
50
Wherein, described duck is that meat duck, egg are with duck or SPF duck.
Further; The invention provides a kind of duck hemorrhagic oaritis inactivated vaccine method for testing efficacy; This method is used duck hemorrhagic oaritis inactivated vaccine immunity 42～560 age in days ducks, carries out 22 weeks after head exempts from and exempts from, and twice immunizing dose only is 0.5 ~ 2ml/; Two exempt from 3 ~ 5 weeks of back attacks poison with the DHOV-HB strain, attacks malicious approach and be oral, intramuscular injection or intravenous injection, and attacking the toxic agent amount is 3 * 10
4ELD
50, attack the poison back and will test duck execution on the 5th～10, cut open inspection, observe the cardinal principle pathology of reproductive system, the malicious protection ratio of attacking of pathology does not take place the reproductive organs of immune group duck is to be judged to vaccine potency more than 70% to be up to the standards; Described cardinal principle pathology shows as that the duck ovarian follicle is hemorrhage, distortion or fallopian tubal atrophy.
Wherein, described duck is that meat duck, egg are with duck or SPF duck.
The present invention has proposed to be used for age in days and the sex that duck hemorrhagic oaritis inactivated vaccine efficacy test uses duck first; The immune time of vaccine potency when check vaccine, attack malicious approach, proposed the qualified standard of vaccine potency test method, criterion and vaccine quality of immunoprotection-cardinal principle pathology judgement.Solved the blank of duck hemorrhagic oaritis inactivated vaccine effect evaluation method and criterion.The method that the present invention confirms is objective, comprehensive, strong operability, can be used for the laboratory and the clinical efficacy evaluation of duck hemorrhagic oaritis inactivated vaccine.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.Under the situation that does not deviate from the present invention's spirit and essence, modification or replacement to the inventive method, step or condition are done all belong to scope of the present invention.
If do not specialize, it is commercially available that used biology, chemical reagent is routine among the embodiment, the conventional means that used technological means is well known to those skilled in the art among the embodiment.Beijing duck used among the embodiment is available from duck hemorrhagic oaritis negative antibody duck field (Nan Kou of Beijing Golden Star Duck Center kind duck field); Sheldrake is available from duck hemorrhagic oaritis negative antibody duck field (dress circle village under four seas town, Yanqing County, Beijing Pearl Spring); The SPF duck is raised to testing required age in days in the SPF chicken isolator by the SPF duck's egg hatching available from Harbin Veterinary Medicine Inst., China Academy of Agriculture's Experimental Animal Center.
Embodiment 1 duck hemorrhagic oaritis inactivated vaccine efficacy test is with the age in days of duck
The Beijing duck of 42,50,100,120,200,300,400,560,600 ages in days is divided into 9 groups, 10 every group, every group of Beijing duck (available from the Nan Kou of Beijing Golden Star Duck Center kind duck field, duck hemorrhagic oaritis antibody is negative) that is same age in days.Test duck oral route is attacked the DHOV-HB strain virus of malicious same dose, contains 3 * 10 approximately
4ELD
50/ only.Attack malicious back 5 days through wing vein collection duck blood, aseptic separation of serum.Serum is inoculated 6 age in days SPF chicken embryos through the yolk bag approach, and 5 pieces/duck, the 0.2ml/ embryo is put 37 ℃ of continuation and hatched to 168h.24h discards with interior dead chicken embryo, and the RT-PCR that calculates interior death of 24～168h and DHOV detects positive chicken embryo number.Having the RT-PCR of 1 piece and above chicken embryo death and DHOV to detect positive person in 5 pieces of chicken embryos of every part of serum is designated as this duck virus and separates positive.According to the neurological susceptibility experimental result, confirm that it is 120 ~ 560 ages in days that duck hemorrhagic oaritis inactivated vaccine efficacy test uses the age in days of duck.
The test stone of susceptible duck or duck embryo
(1) 5 of susceptible ducks are from without the immunity of duck hemorrhagic oaritis inactivated vaccine or the popular history duck of duck hemorrhagic oaritis crowd's duck is arranged, and strain is used in the check of 100 times of dilutions of every oral 1.0 milliliters of aseptic 0.5% lactoalbumin hydrolysate Hank ' s liquid of duck.Attacked poison back 5 days, blood sampling, separation of serum carries out virus to be separated.The virus separation rate should at least 4/5 positive.
(2) 10 pieces of susceptible embryos are from without immunity of duck hemorrhagic oaritis inactivated vaccine or the kind egg that has duck hemorrhagic oaritis pestilence history kind duck crowd to be produced, 37 ℃ of hatching to 10 ages in days.Inoculate the kind poison of any generation of 500 times of dilutions of 0.5% aseptic lactoalbumin hydrolysate Hank ' s liquid (LH) through allantoic cavity, every embryo 0.1ml puts and continues hatching under 36～37 ℃ of conditions.The duck embryo should be in inoculation back death in 24～96 hours at least 9/10.It is systemic bleeding that dead duck embryo is observed pathology.
Embodiment 2 duck hemorrhagic oaritis inactivated vaccine effect evaluations are with the sex of duck
The Beijing duck oral route of 80 140 age in days male and female half and half is attacked the DHOV-HB strain virus of malicious same dose, contain 3 * 10 approximately
4ELD
50/ only.Attack malicious back 1～8 day through wing vein collection duck blood, 10/day, aseptic separation of serum.Serum is inoculated 6 age in days SPF chicken embryos through the yolk bag approach, and 5 pieces/duck (5 pieces of chicken embryos of every duck serum inoculation), the 0.2ml/ embryo is put 37 ℃ of continuation and is hatched to 168h.24h discards with interior dead chicken embryo, calculates death and the positive chicken embryo of DHOV number in 24～168h.Having 1 piece and above chicken embryo death and DHOV positive person to be designated as this duck virus in 5 pieces of chicken embryos of every part of serum separates positive.Female Beijing duck virus is separated positive rate apparently higher than male Beijing duck.The positive rate of the virus infections chicken embryo that the poison of attacking against each other afterwards occurs behind the male and female duck serum inoculated into chick embryo every day carries out the paired t check of two samples, and the P value is less than 0.05, and there were significant differences aspect neurological susceptibility for sex.
Immunoprotection-cardinal principle pathology the method (1) of embodiment 3 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 200 age in days Beijing ducks of duck hemorrhagic oaritis inactivated vaccine (lot number 2012001, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 0.5ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 5 weeks, and attacking poison strain is the DHOV-HB strain, and intramuscular injection path is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the Beijing duck of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 10th; Cut open inspection; Atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck, the test duck that ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is not all normally had pathology is judged to protection.The malicious protection ratio of attacking of immune group duck is 80% in the present embodiment, and the incidence of disease of non-immune control group duck is 80%, explains that this batch vaccine potency is up to the standards.
Immunoprotection-cardinal principle pathology the method (2) of embodiment 4 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 140 age in days Beijing ducks of duck hemorrhagic oaritis inactivated vaccine (lot number 2012002, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 1ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 4 weeks, and attacking poison strain is the DHOV-HB strain, and the intravenous injection approach is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the Beijing duck of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 9th; Cut open inspection; Atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck, the test duck that ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is not all normally had pathology is judged to protection.The malicious protection ratio of attacking of immune group duck is 70% in the present embodiment, and the incidence of disease of non-immune control group duck is 80%, explains that this batch vaccine potency is up to the standards.
Immunoprotection-cardinal principle pathology the method (3) of embodiment 5 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 300 age in days Beijing ducks of duck hemorrhagic oaritis inactivated vaccine (lot number 2012003, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 1.5ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 3 weeks, and attacking poison strain is the DHOV-HB strain, and the oral infection approach is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the Beijing duck of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 8th; Cut open inspection, atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck; Ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is all normal, and the test duck of no pathology is judged to protection.The malicious protection ratio of attacking of immune group duck is 90% in the present embodiment, and the incidence of disease of non-immune control group duck is 100%, explains that this batch vaccine potency is up to the standards.
Immunoprotection-cardinal principle pathology the method (4) of embodiment 6 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 400 age in days Beijing ducks of duck hemorrhagic oaritis inactivated vaccine (lot number 2012004, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 2ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 5 weeks, and attacking poison strain is the DHOV-HB strain, and intramuscular injection path is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the Beijing duck of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 7th; Cut open inspection; Atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck, ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is all tested duck normally be judged to protection.The malicious protection ratio of attacking of immune group duck is 100% in the present embodiment, and the incidence of disease of non-immune control group duck is 90%, explains that this batch vaccine potency is up to the standards.
Immunoprotection-cardinal principle the pathology (5) of embodiment 7 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 560 age in days Beijing ducks of duck hemorrhagic oaritis inactivated vaccine (lot number 2012005, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 1ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 4 weeks, and attacking poison strain is the DHOV-HB strain, and intramuscular injection path is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the Beijing duck of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 7th; Cut open inspection; Atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck, the test duck that ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is not all normally had pathology is judged to protection.The malicious protection ratio of attacking of immune group duck is 80% in the present embodiment, and the incidence of disease of non-immune control group duck is 100%, explains that this batch vaccine potency is up to the standards.
Immunoprotection-cardinal principle pathology the method (6) of embodiment 8 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 120 age in days Beijing ducks of duck hemorrhagic oaritis inactivated vaccine (lot number 2012006, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 1.5ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 4 weeks, and attacking poison strain is the DHOV-HB strain, and oral route is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the Beijing duck of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 6th; Cut open inspection; Atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck, the test duck that ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is not all normally had pathology is judged to protection.The malicious protection ratio of attacking of immune group duck is 100% in the present embodiment, and the incidence of disease of non-immune control group duck is 90%, explains that this batch vaccine potency is up to the standards.
Immunoprotection-cardinal principle pathology the method (7) of embodiment 9 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 140 age in days sheldrakes of duck hemorrhagic oaritis inactivated vaccine (lot number 2012001, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 1.0ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 4 weeks, and attacking poison strain is the DHOV-HB strain, and oral route is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the sheldrake of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 6th; Cut open inspection; Atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck, the test duck that ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is not all normally had pathology is judged to protection.The malicious protection ratio of attacking of immune group duck is 70% in the present embodiment, and the incidence of disease of non-immune control group duck is 90%, explains that this batch vaccine potency is up to the standards.
Immunoprotection-cardinal principle pathology the method (8) of embodiment 10 duck hemorrhagic oaritis inactivated vaccine effect evaluations
Use 10 of the immune 120 age in days SPF ducks of duck hemorrhagic oaritis inactivated vaccine (lot number 2012001, Beijing City Agriculture and Forestry Institute Raise Livestockv Veterinarian Institute and Ruipu's development), male and female half and half are as the vaccine immunity group.The immunizing dose of vaccine immunity group and method are: first immune seedling dosage is 1.0ml/, the chest muscle injection.Head exempts from 2 weeks of back, exempts from identical approach two according to same dose, and two exempt from the back attacked poison in 4 weeks, and attacking poison strain is the DHOV-HB strain, and oral route is attacked poison, and every duck is attacked the toxic agent amount and is about 3 * 10
4ELD
50Establish simultaneously do not infect DHOV and not the SPF duck of immune duck hemorrhagic oaritis inactivated vaccine susceptible (10, male and female half and half are with the same age in days of immune group duck) attack poison for control group.Attack the poison back and put to death the experiment duck on the 5th day; Cut open inspection; Atrophy is appearred in ovary or fallopian tubal (testis or fallopian tubal), or ovarian follicle is out of shape and hemorrhage test duck is judged to the morbidity duck, the test duck that ovary, ovarian follicle and fallopian tubal (testis and vas deferens) is not all normally had pathology is judged to protection.The malicious protection ratio of attacking of immune group duck is 80% in the present embodiment, and the incidence of disease of non-immune control group duck is 100%, explains that this batch vaccine potency is up to the standards.
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from know-why of the present invention; Can also make some improvement and retouching, these improvement and retouching also should be regarded as protection scope of the present invention.
Claims (8)
1. duck hemorrhagic oaritis inactivated vaccine method for testing efficacy; It is characterized in that use duck hemorrhagic oaritis inactivated vaccine immune duck, immunity is 2 times altogether; Carry out 22 weeks after head exempts from and exempt from, two exempt from 3 ~ 5 weeks of back attacks poison with duck hemorrhagic oaritis virus to immune duck; Attack the poison back and will test duck execution on the 5th～10, cut open inspection, observe the cardinal principle pathology of reproductive system, the malicious protection ratio of attacking of pathology does not take place the reproductive organs of immune group duck is to be judged to vaccine potency more than 70% to be up to the standards; Described cardinal principle pathology shows as duck that the duck ovarian follicle is hemorrhage, distortion or fallopian tubal atrophy, shows as testis or atrophy of vas deferens male duck.
2. the method for claim 1 is characterized in that, it is 120～560 ages in days that efficacy test uses the age in days of duck.
3. the method for claim 1 is characterized in that, it is female that efficacy test uses the sex of duck.
4. the method for claim 1 is characterized in that, the dosage of said 2 immunity be 0.5ml/ only～2ml/ only.
5. the method for claim 1 is characterized in that, the described poison of attacking uses duck hemorrhagic oaritis virus to be the DHOV-HB strain, and attacking the toxic agent amount is 3 * 10
4ELD
50
6. like the arbitrary said duck hemorrhagic oaritis inactivated vaccine method for testing efficacy of claim 1-5, it is characterized in that described duck is that meat duck, egg are with duck or SPF duck.
7. a duck hemorrhagic oaritis inactivated vaccine method for testing efficacy is characterized in that, uses duck hemorrhagic oaritis inactivated vaccine immunity 120～560 age in days ducks, carries out 22 weeks after head exempts from and exempts from, and twice immunizing dose only is 0.5 ~ 2ml/; Two exempt from 3 ~ 5 weeks of back attacks poison with the DHOV-HB strain, attacks malicious approach and be oral, intramuscular injection or intravenous injection, and attacking the toxic agent amount is 3 * 10
4ELD
50, attack the poison back and will test duck execution on the 5th～10, cut open inspection, observe the cardinal principle pathology of reproductive system, the malicious protection ratio of attacking of pathology does not take place the reproductive organs of immune group duck is to be judged to vaccine potency more than 70% to be up to the standards; Described cardinal principle pathology shows as that the duck ovarian follicle is hemorrhage, distortion or fallopian tubal atrophy.
8. like the said duck hemorrhagic of claim 7 oaritis inactivated vaccine method for testing efficacy, it is characterized in that described duck is that meat duck, egg are with duck or SPF duck.
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