CN104130981A - Application of avian infectious bronchitis virus vaccine strain in preparation of inactivated vaccine - Google Patents
Application of avian infectious bronchitis virus vaccine strain in preparation of inactivated vaccine Download PDFInfo
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Abstract
The invention discloses an application of avian infectious bronchitis virus vaccine strain in the preparation of inactivated vaccine. The separated wild virulent strain of infectious bronchitis virus is subjected to chick embryo adaptability domestication so as to obtain an infectious bronchitis virus vaccine strain, which can well adapt to the chick embryo, maintains good hereditary characters and immunogenicity, and is named as ck/CH/LLN101135. Then the obtained infectious bronchitis virus vaccine strain is inactivated and subjected to immune efficacy evaluation experiments. The experiment results show that the infectious bronchitis inactivated vaccine prepared from the vaccine strain can trigger the organism to generate a good immune response, can prominently improve the motive immune protective effectiveness of chicken at the same time, and effectively protects chicken from being affected by infectious bronchitis virus. The infectious bronchitis virus vaccine strain can be used to prepare single vaccine or combined vaccine of avian infectious bronchitis.
Description
Technical field
The present invention relates to virus vaccine strain, relate in particular to the virus vaccine strain of chicken infectious bronchitis and in the application of preparing in inactivated vaccine, the invention belongs to the prevention and control field of chicken infectious bronchitis.
Background technology
Chicken infectious bronchitis (Avian Infectious Bronchitis, IB) be by avian infectious bronchitis virus (Infectious Bronchitis Virus, IBV) one that causes is acute, height contagiousness, viral respiratory disease, it is characterized in that: sick chicken coughs, sneezes, tracheae rale; Kidney enlargement, pale, ureterectasia, has a large amount of urate deposition, outward appearance to present typically " piebald kidney ".Chicken infectious bronchitis is generally popular in countries in the world since nineteen thirty, has caused larger financial loss.Along with viral propagation and Genetic evolution, there is multiple serotype in IBV at present.
The popular IBV in most of China area at present, and all have the pathogenic IBV of New Kidney popular in Chinese vaccinated flock and nonimmune chicken group.This disease is as one of serious infectious diseases of serious harm China poultry husbandry, and many new variants generally appear in this cause of disease in recent years, has brought new threat to China's aquaculture.In order effectively to prevent and treat this disease, be necessary to study new generation vaccine, present stage, its conventional vaccine was deactivation vaccine or the weak malicious seedling H120 of its chicken embryo adaptation and the H52 of the widely used M41 serotype in countries in the world.But because IBV serotype is numerous at present; existing vaccine all can only provide part protection or unprotect; and IB attenuated vaccine causes weak degree and is difficult to grasp; can there is certain pathogenic and transmission capacity; even may in the body infecting, cause lasting latent infection; become the main body of sudden change or the donor of restructuring variation, add that the conditional request such as transport, storage and use of vaccine is higher, make to cause the weak malicious vaccine of living and all can not fundamentally thoroughly control the popular of IB in theory with in practice.
Inactivated vaccine security is good, does not exist to disseminate cause of disease and virulence is returned strong problem, and can excite good humoral immune reaction.The M41 inactivated vaccine of current domestic extensive application has been brought into play vital role in production in the past, but, complicated along with China IB epidemic status, this inactivated vaccine strain serotype can not be mated completely with new epidemic isolates, causes infectious bronchitis generally to occur in vaccinated flock.
Therefore, development is extremely urgent with corresponding living vaccine integrated application to China's epidemic isolates inactivated vaccine with strong points.The present invention is just based on above situation, utilize existing Research foundation, the novel infectious bronchitis vaccines strain of screening and cultivating is prepared into safety, efficient new generation vaccine, research shows, this inactivated vaccine can effectively be resisted the attack of Major Epidemic infectious bronchitis street strain of current China, for effective control chicken infectious bronchitis epidemic disease continue break out with popular significant.
Summary of the invention
Technical problem to be solved by this invention be overcome existing inactivated vaccine for avian infectious bronchitis strain M41 (viral spike protein (S1) genetic evolution close fasten with Massachusetts close, those skilled in the art are summarized as Mass type chicken infectious bronchitis strain) can not effectively anti-chicken infectious bronchitis processed (Avian Infectious Bronchitis, IB) attack of epidemic isolates, the defect such as the prevention effect of IB is undesirable, the new infectious bronchitis vaccines strain of one strain is provided, and (viral spike protein (S1) genetic evolution pass is fastened close with LX4 strain or QX strain, those skilled in the art are summarized as LX4 type (QX sample) strain), this vaccine strain can effectively be resisted the attack of Major Epidemic infectious bronchitis street strain of current China, good immune protection effectiveness can be provided.
Technical problem to be solved by this invention realizes by following technological approaches:
The present invention will filter out and have Typical Representative and after deactivation, have good antigenic LX4 type (QX sample) infectious bronchitis vaccines strain from the popular infectious bronchitis epidemic isolates of China, its morphological observation: virus strain allantoic fluid can be seen diameter through the crown poison for 80-120nm after centrifugal, phospho-wolframic acid negative staining under electric ytterbium, poison virus particle is polymorphism, most is circular, have cyst membrane, there is the coronal process that is loose, evenly distributed on surface.
A kind of infectious bronchitis vaccines strain of the present invention, called after ck/CH/LLN101135, Classification And Nomenclature is infectious bronchitis virus Infectious bronchitis virus, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences, preservation date is on February 21st, 2014, and its microbial preservation number is: CGMCC No.8790.
Its viral spike protein subunit S1 gene order evolutionary relationship of infectious bronchitis vaccine strain ck/CH/LLN101135 strain involved in the present invention is classified as shown in Figure 1.S1 gene is the main protective gene of infectious bronchitis virus; can stimulate body to produce neutralizing antibody; S1 gene is also the gene the most easily morphing in viral evolutionary process; and its hypervariable region is mainly positioned at S1 gene; therefore the present invention mainly shows the S1 gene order that the present invention relates to strain; the aminoacid sequence of the spike protein S1 subunit that vaccine strain of the present invention has, as shown in SEQ ID NO:1, is encoded by SEQ ID NO:2.Spike protein subunit S1 gene has more than 95% homology compared with sequence shown in SEQ ID NO:2; and there is the virus strain of same or similar immune efficacy or the S1 transgenation strain of vaccine strain ck/CH/LLN101135 of the present invention strain with avian infectious bronchitis virus vaccine strain ck/CH/LLN101135 of the present invention strain, as long as the spike protein subunit S1 gene after sudden change has within more than 95% homology just still drops on protection scope of the present invention compared with sequence shown in SEQ ID NO:2.LX4 type (QX sample) vaccine strain the present invention relates to can continue propagation and evolve in chicken embryo or chicken body, the strain that those skilled in the art can cultivate by separation contrasts the S1 gene of S1 gene and LX4 type (QX sample) strain, can find that LX4 type (QX sample) vaccine strain the present invention relates to closes the feature of fastening in S1 genetic evolution.
By after a kind of infectious bronchitis vaccines strain ck/CH/LLN101135 strain deactivation of the present invention, carry out immune efficacy evaluation experimental, result shows, ck/CH/LLN101135 vaccine strain of the present invention still can keep good immunogenicity after inactivator inactivation treatment, be inoculated in the significantly neutralizing antibody level of lift pins to LX4 type (QX sample) virus of chicken through Mass type infectious bronchitis living vaccine H120 fundamental immunity, wherein the serum NAT of 10 ck/CH/LLN101135 strain inactivated vaccine booster immunization chickens is no less than 6 compared with the high 2 times of above chicken quantity of the serum NAT of 10 Mass type living vaccine H120 fundamental immunity chickens, and the serum NAT of ck/CH/LLN101135 strain inactivated vaccine booster immunization chicken all contrasts the average serum NAT of chicken higher than Mass type vaccine strain H120 fundamental immunity, illustrate that the ck/CH/LLN101135 strain vaccine strain the present invention relates to has good lifting effect to chicken infectious bronchitis protection antibody.Simultaneously, ck/CH/LLN101135 strain inactivated vaccine booster immunization chicken and Mass type vaccine strain H120 fundamental immunity chicken are with after LX4 type (QX sample) strong virus attack, serum neutralizing antibody (for LX4 type (QX sample) strain) titre of ck/CH/LLN101135 strain inactivated vaccine booster immunization chicken and the serum NAT difference of Mass type living vaccine H120 fundamental immunity chicken are remarkable equally, wherein the serum NAT of 10 ck/CH/LLN101135 strain inactivated vaccine booster immunization chickens is no less than 7 compared with the high 2 times of above chicken quantity of the serum NAT of 10 Mass type living vaccine H120 fundamental immunity chickens, and the serum NAT of ck/CH/LLN101135 strain inactivated vaccine booster immunization chicken all contrasts the average serum NAT of chicken higher than Mass type vaccine strain H120 fundamental immunity, illustrate that the ck/CH/LLN101135 strain vaccine strain the present invention relates to has good lifting effect to chicken infectious bronchitis protection antibody.Safety testing result shows, LX4 type (QX sample) inactivated vaccine for avian infectious bronchitis the present invention relates to is to chick and kind of the chicken safety of laying eggs, and without side reaction, security is good.
Another technical problem to be solved by this invention is to provide described avian infectious bronchitis virus vaccine strain in the application of preparing in inactivated vaccine for avian infectious bronchitis.
Another technical problem to be solved by this invention is to provide the vaccine composition of a kind of control chicken infectious bronchitis (LX4 type (QX sample)), and on its avian infectious bronchitis virus vaccine strain of the present invention (called after ck/CH/LLN101135 strain) after by deactivation and pharmacology, acceptable immunological adjuvant forms.
The vaccine composition of a kind of control chicken infectious bronchitis of the present invention (LX4 type (QX sample)) can prepare with reference to following methods: (1) is cultivated propagation by the vaccine strain ck/CH/LLN101135 strain the present invention relates to and obtained production seed culture of viruses; Production is inoculated in chick embryo allantoic cavity with seed culture of viruses, cultivates, gather in the crops viral allantoic fluid; (2) measure qualified viral allantoic fluid through steriling test and viral level and add inactivator to carry out inactivation treatment, prepare antigen; (3) antigen being up to the standards through deactivation and adjuvant carry out emulsification, are prepared into LX4 type (QX sample) infectious bronchitis inactivated vaccine.
The using method of LX4 type (QX sample) inactivated vaccine for avian infectious bronchitis the present invention relates to:
(1) route of inoculation: muscle or subcutaneous vaccination.
(2) dosage of inoculation: 1 plumage part (0.3-0.8ml).
The immunity of LX4 type of the present invention (QX sample) inactivated vaccine for avian infectious bronchitis produces immunizing power in latter 20 days, and the primary immune response extended period is 4 months.LX4 type of the present invention (QX sample) strain, except using as single seedling, also can be used for preparation connection seedling etc.
Brief description of the drawings
Fig. 1 ck/CH/LLN101135 strain vaccine of the present invention strain and the Reference strains genetic evolution graph of a relation based on viral spike protein S1 gene.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment are only exemplary, scope of the present invention are not formed to any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these amendments and replacement all fall within the scope of protection of the present invention.
The isolation identification of the avian infectious segmental bronchus strain of embodiment 1 ck/CH/LLN101135
1, materials and methods
1.1 terrain samples separate with virus: IBV is carried out in lasting epidemiological surveillance process at us, the chicken group of plant that doubtful IB infects from China various places collects kidney sample, the early clinic symptom of morbidity chicken performance respiratory tract disease, comprises and breathes, coughs, sneezes and tracheae rale.Dead chicken is cutd open inspection, visible trachitis, ephritis and proventriculitis in various degree, the death of laying hen group main manifestations, egg drop reduction and produce lopsided egg.The sample source that this research is collected is crossed the chicken group of Mass type commercialization attenuated vaccine in immunity.
When virus separates, sample is ground to the PBS damping fluid that is suspended in 0.1%, supernatant is filtered by the filtering membrane of 0.22 μ m after the centrifugal 10min of 1500g in 4 DEG C, be inoculated in 9-11 age in days SPF chick embryo allantoic cavity, after inoculation, embryo being placed in to 37 DEG C hatches, every day, 3-5 blind passage of process was until there is the characteristic pathologies such as dwarf embryo, dysplasia, curling embryo for 2-7 days in chicken embryo culture according to embryo.
1.2 electron microscopic observations: occur typical cytopathic after sample inoculated into chick embryo, after being processed, allantoic fluid carries out electron microscopic observation, by 1.5ml allantoic fluid after the centrifugal 30min of low speed 1500g, supernatant is with the centrifugal 30min of 12000g, get supernatant,, dye and observe under Electronic Speculum by negative staining resuspended the precipitation after centrifugal with a little deionized water.
1.3 IBV vaccine and strains: commercialization Mass type vaccine H120 is for Immunization protection test, and eye droppings approach is inoculated to specifications.LX4 type (QX sample) the representative strains ck/CH/LLN101135 strain that this research separates is for immunoprotection evaluation test, and by Reed-Muench method measurement and calculation virus titer, inoculation chicken is front by viral dilution to 10
4.8eID
50/ 100 microlitres, a 100 microlitres/chicken inoculates.
Clone and the sequencing of 1.4 IBV strain isolated S1 genes: the S1 gene nucleotide series of the MEGALIGN program of utilizing DNAStar software to strain isolated and Reference strains and the aminoacid sequence of derivation carry out contrast and analysis, utilize the Clustal V method of DNAStar software to carry out phylogenetic analysis to S1 gene nucleotide series, the Reference strains that this research is selected is for phylogenetic analysis, and these strains have represented China different time and the strain of different areas and the LX4 type of part Foreign Epidemic (QX sample) IBV strain during 1997-2011.
1.5 experimentation on animals designs: represent that taking strain LX4 type (QX sample) IBV strain ck/CH/LLN101135 strain is as example, 60 1 age in days SPF chickens are divided into 4 groups, raise respectively in independent negative pressure isolator, 1-2 organizes every group of 20 chickens, and the 3rd and 4 groups every group 10.1st, inoculate H120 vaccine at 1 age in days according to vaccine specification sheets for 3 groups; 2nd, 4 groups of blank allantoic fluids that chicken inoculation is aseptic; Wherein the 1st group and the 2nd group collunarium in the time of 20 age in days is attacked LX4 type (QX sample) IBV and is represented strain ck/CH/LLN101135 strain, the 3rd group and the 4th group respectively with identical by way of the aseptic blank allantoic fluid of inoculation.
1.6 experimentation on animals sample collectings: the 1st, get 10 chickens and catch and kill attacking every group of latter 5 days of poison for 2 groups, get respectively 5 and catch and kill for the 3rd group and the 4th group, tracheae and the kidney of chicken catched and killed in collection, individual curing is placed in viral parting liquid (50% glycerine of 300 microlitres respectively, 50%PBS), be stored under-20 DEG C of conditions and separate in order to virus, every group of residue chicken carries out clinical observation, observe and catch and kill and cut open inspection after 30 days, immunity chicken is after immunity 3, 6, 9, 12, blood sampling in 15 and 18 days, separation of serum is also measured serum antibody, attack malicious chicken and attack poison rear 3, 6, 9, same blood sampling in 12 and 15 days, separation of serum carries out antibody test.
In 1.7 experimentation on animals samples, the separation of virus is identified with RT-PCR: attack every the chicken sample gathering after poison and all separate for virus, each sample adds 0.1%PBS in 10% ratio after grinding, first with 1500g at 4 DEG C of centrifugal 10min, supernatant is inoculated in 9-11 age in days SPF chick embryo allantoic cavity after the membrane filtration of 0.22 μ m, 0.2ml/ embryo, 3 pieces of embryos of each sample inoculation, the rear chicken embryo of inoculation is put 37 DEG C and is continued to hatch, every day is according to embryo, every kind of sample is inoculated latter 72 hours and is got wherein 2 pieces of embryo allantoic liquids and carries out RT-PCR amplification, residue chicken embryo is observed idiosome characteristic pathology after one week, as whether there is dwarf embryo, arrested development and curling embryo.Through 1-3 for blind passage, chicken embryo death or occur that the typical characteristic pathology of IB is judged to the sample IBV positive.
In order to ensure accurately determining of judgement, the allantoic fluid of inoculated into chick embryo is carried out to RT-PCR amplification qualification, getting 200 microlitre allantoic fluids increases for RT-PCR, extracted total RNA, utilize N (-) primer to carry out reverse transcription, primer N (-) and N (+) is used for the fragment of the approximately 1600bp size of amplification including most of N gene and 3 ' UTR subregion, and the PCR product of amplification is analyzed by 1% agarose gel electrophoresis.
1.8 antibody tests: utilize the commercialization ELISA test kit of IDEXX company to detect the serum sample of collection, and calculate the positive ratio of serum antibody according to test kit specification sheets.
2 results
2.1 viruses detect and separating resulting:
The sample infecting from doubtful IB, be separated to IBV virus, the allantoic fluid of different generations after inoculated into chick embryo is carried out to electron microscopic observation, all strain isolateds all have the representative configuration feature of coronavirus particle, and exist without other exogenous viruses in sample.
The wild virulent strain of infectious bronchitis virus that the present invention obtains separation carries out chicken embryo and adapts to domestication, obtain and highly adapt to chicken embryo and keep good hereditary property and immunogenic infectious Bronchitis Virus Vaccine strain, called after ck/CH/LLN101135 strain, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address is in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City institute of microbiology of the Chinese Academy of Sciences, preservation date is on February 21st, 2014, and its microbial preservation number is: CGMCC No.8790.
The phylogenetic analysis of 2.2 IBV isolated strains:
By the phylogenetic analysis of the S1 gene order to the IBV strain ck/CH/LLN101135 strain separating and Reference strains, show that isolated in China strain can be divided into 5 different gene groups or genotype, as shown in Figure 1, the IBV strain ck/CH/LLN101135 strain separating is with openly Reference strains LX4 and QX strain belong to a genotype, this group of strains are China's Major Epidemic strain, mainly comprise the strain that China separates during 1997-2009.And LX4 type (QX sample) strain of finding isolated in China by contrast is higher with external strain similarity, show that the LX4 type that country variant is popular (QX sample) strain source is similar.
2.3 experimentation on animals clinical observation result:
Test chicken is with within after virus attack 3 days, starting to occur clinical symptom, and performance feather is fluffy and disorderly, do not eat or few food, but all do not show obvious respiratory symptom, and contrast chicken is acted normally, and attacks lassitude of malicious chicken, poly-heap.Attacking latter 5 days chicken groups of poison all falls ill, attack the rear 5-7 days of poison, wherein the attack of ck/CH/LLN101135 strain virus is only organized dead 2-5, dead chicken is dissected and finds all to occur kidney enlargement, pale, ureter and kidney all have urate deposition, show that ck/CH/LLN101135 strain strain is for causing nephropathy modification, and also find to attack the rear dead SPF chicken cecal tonsil of poison all has hemorrhage simultaneously.But all do not observe obvious pathology at glandular stomach and uterine tube, period of disease is for attacking the rear 3-10 days of poison.
2.4 Mass type vaccine H120 are to separating malicious protectiveness:
Attacking all discovery death of chicken after H120 immunity although separate poison, there is clinical symptom in partial immunity chicken, illustrates that H120 vaccine can not provide protection completely to the ck/CH/LLN101135 strain virus separating.Attack latter 5 days collected specimens of poison and carry out virusology detection, the results are shown in Table 1.Tracheae and the kidney of visible nonimmune contrast chicken are all separated to virus; and the chicken of all immune H120 has all been separated to too virus in tracheae; in the kidney of most of immune chickens, be also separated to virus, illustrate and can not attack the protection that provide kidney and tracheae to the ck/CH/LLN101135 strain strain of isolated in China with the immunity of Mass type vaccine virus.
2.5 results of serological detections:
To immune chicken with attack malicious chicken and detect at the serum of different time collection, the results are shown in Table 1.Every group of immunity chicken all has 1 chicken antibody to fail to turn sun for latter 6 days in H120 immunity, the antibody of latter 15 days all H120 immunity chickens of immunity all turns positive, from the result of measuring, the antibody of wild virus infection turns sun will turn the positive time early than antibody after vaccine virus immunity, and it is stronger that this also further reacts ck/CH/LLN101135 strain strain immunogenicity.
The rear serological reaction of poison and viral separating resulting are attacked in table 1 ck/CH/LLN101135 strain
3 conclusions:
Be separated to different LX4 type (QX sample) infectious bronchitis virus from China different areas and represent strain ck/CH/LLN101135 strain; for typical LX4 type (QX sample) represents strain; this type strain virulence is strong, and the Mass type vaccine strain that was in the past applied to IBV prevention and control can not effectively be protected the attack of this type strain ck/CH/LLN101135 strain.Therefore, LX4 type (QX sample) the infectious bronchitis series vaccine of research based on China's Major Epidemic is all significant for the IB preventing control program of China or other countries.
The immune efficacy experiment of embodiment 2 infectious bronchitis inactivated vaccine of the present invention strain ck/CH/LLN101135 strain
1, materials and methods
1.1 fundamental immunity vaccines
Mass type infectious bronchitis inactivated vaccine (H120 strain), purchased from Harbin Wei Ke biotechnology development company, vaccine preservation condition and validity period :-15 DEG C of following preservations, validity period is 12 months.Press label mark plumage part with drawing vaccine after sterile saline dilution, every chicken eye droppings collunarium is inoculated 1 plumage part.
The propagation of 1.2 viral LX4 type (QX sample) infectious bronchitis virus ck/CH/LLN101135 strains
Use 10 age in days SPF chicken embryos to carry out 10000 times of dilutions to ck/CH/LLN101135 strain kind poison, every piece of embryo is inoculated by allantoic cavity, every embryonic breeding kind 100 μ L, culture condition is: hatch 72h from 37 DEG C, after 72h, chicken embryo is taken out, observe chicken embryo pathology situation, aseptic collection chick embryo allantoic liquid.
1.3 virus titers are measured
Get the ck/CH/LLN101135 chick embryo allantois virus liquid of propagation, make successively respectively 10 times of serial dilutions; Select 4 extension rates (10
4～10
7or 10
5～10
8) viral suspension inoculate respectively the SPF chicken embryo of 5 piece of 10 age in days, every piece of egg inoculation 100 μ L, put chicken embryo in 37 DEG C of incubators and hatch 1 week, every day is according to embryo, discard dead embryo within 24h, record infection number and the existence number of chicken embryo in 1 week, calculate EID by Reed-Muench method
50.With>=10
6eID
50/ 100
μl allantoic fluid is qualified.-80oC saves backup.
1.4 steriling test
Ck/CH/LLN101135 strain virus allantoic fluid is carried out to steriling test by existing " People's Republic of China's veterinary drug allusion quotation " annex.
The deactivation of 1.5 ck/CH/LLN101135 virus liquids
By the virus liquid being up to the standards, add inactivator (formaldehyde) deactivation, making its final concentration is 0.2%, fully shakes up immediately.Then put deactivation under 37 DEG C of conditions, after 24 hours deactivation complete, the virus liquid after deactivation is put 2～8 DEG C and is saved backup.
The deactivation inspection of 1.6 ck/CH/LLN101135 virus liquids
Get the allantoic fluid after deactivation, 10 pieces of inoculation 10 age in days SPF chicken embryos in allantoic cavity, every embryo 0.2ml, puts under 37 DEG C of conditions and hatches, and discards dead chicken embryo in 24 hours, observe 168 hours, the non-specific death of chicken embryo should be no more than 2, checks one by one chicken embryo, should not occur dehydration, rolls up, the characteristic lesion such as dwarf embryo, and 1 generation of blind passage, should be without dehydration, roll up, the characteristic lesion such as dwarf embryo thinks that deactivation is complete.
The preparation of 1.7 oil emulsion inactivated vaccines
Utilize respectively injection white oil and span-80 and aluminum stearate to prepare oil phase, ck/CH/LLN101135 deactivation allantoic fluid and tween-80 are prepared water, and add sanitas to adopt emulsifying device to carry out fully emulsified.Qualified emulsification vaccine is sub-packed in sterilizing vaccine bottle, and 2～8 DEG C of preservations, in order to inspection.
The effect evaluation of 1.8 infectious bronchitis oil emulsion inactivated vaccines
40 1 age in days SPF chickens are divided into 2 groups (20 every group) at random, in negative pressure isolator, raise and whole immune Mass type infectious bronchitis living vaccines (H120 strain), and 1 plumage part/only.Wherein one group of chick is exempted from latter 20 days subcutaneous inoculation inoculation LX4 type (QX sample) infectious bronchitis inactivated vaccines in H120 head (ck/CH/LLN101135 strain, wherein every plumage part vaccine is>=10 at deactivation provirus content
6eID
50), every subcutaneous vaccination 0.5ml.Another group chick is as H120 living vaccine fundamental immunity control group.Booster immunization group is respectively got 10 chickens blood samplings separation of serum in two groups of immunity rear 20 days, and application serum neutralization test method detects the serum NAT of every chicken.2 groups of residue chick are carried out to eye droppings and collunarium attack with strong malicious ck/CH/LLN101135 simultaneously, attack toxic agent amount for every and be no less than 10
5.0eID
50attack poison and within latter 5 days, catch and kill booster immunization group and fundamental immunity control group chicken, blood sampling separation of serum respectively, gather kidney, the tracheae of every chicken simultaneously, the serum gathering is measured respectively for the neutralization of ck/CH/LLN101135 strain and tired, measure the IBV target organ tracheae and the kidney that gather simultaneously, carry out virus isolated rate mensuration.Booster immunization group serum antibody titer and fundamental immunity contrast the difference between chicken, the effect of evaluation infectious bronchitis inactivated vaccine before attacking poison by contrast and after attacking poison.
2 results
The virus titer measurement result of 2.1 ck/CH/LLN101135 strain virus liquid
In the allantoic fluid of propagation, viral level is 10
6.2eID
50/ 0.1ml.
2.2 steriling test results
The chick embryo allantoic liquid of virus of proliferation is through inspection no bacteria pollution.
2.3 LX4 types (QX sample) infectious bronchitis inactivated vaccine immune efficacy evaluation result
After Mass type living vaccine head exempts from, after LX4 type (QX sample) infectious bronchitis inactivated vaccine (ck/CH/LLN101135) booster immunization, 10 chicken serum NATs of 20 days reach 8 compared with the high more than 2 times chicken quantity of highest serum NAT of 10 fundamental immunity contrast chickens, and the serum NAT of inactivated vaccine booster immunization chicken all contrasts the average serum NAT of chicken higher than fundamental immunity.Simultaneously, 10 chicken serum NATs of attacking poison after inactivated vaccine booster immunization are attacked the high 2 times of above chicken quantity of malicious fundamental immunity contrast chicken serum NAT compared with 10 and are reached 9, and the serum NAT of inactivated vaccine booster immunization chicken is all higher than the average serum NAT of fundamental immunity contrast chicken.Attack malicious chicken target organ virus separation detection result and show, booster immunization chicken tracheae separates positive rate than the low 20-30% of fundamental immunity control group (2-3/10) with virus in kidney.The results are shown in Table 2.
Table 2 booster immunization group and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
3. conclusion (of pressure testing)
After Mass type living vaccine head exempts from, can promote virucidin's level by effective stimulus body with LX4 type (QX sample) infectious bronchitis inactivated vaccine booster immunization, the validity of infectious bronchitis inactivated vaccine has been described.
The safety experiment of embodiment 3 LX4 type of the present invention (QX sample) infectious bronchitis inactivated vaccine (ck/CH/LLN101135)
1 experiment material
1.1 test vaccines:
Prepared 4 batches of vaccines according to the method for embodiment 2, the lot number of 4 batches is respectively: 2012001,2012002,2012003,2012004,500 plumage part/bottles.Vaccine preservation condition and validity period: 2～8 DEG C of preservations, validity period is 12 months.Draw vaccine by label mark plumage part with sterilizing syringe, under every cock skin or intramuscular inoculation.
1.2 laboratory animal:
1 week age, SPF chicken was purchased from Harbin Veterinary Medicine Inst., China Academy of Agriculture's Experimental Animal Center.
2 experimental techniques
2.1 SPF chicken proof tests
2.1.1 single dose inoculation test
Single dose inoculation is 30 1 ages in days and 30 6 Japanese instar chicklings for proof test, and every batch of vaccine of 2012001 and 2012002 batches of vaccines uses 10 1 Japanese instar chicklings, separately establishes 10 of control group chickens; Every batch of vaccine of 2012003 and 2012004 batches of vaccines uses 10 6 Japanese instar chicklings, separately establish 10 of control group chickens, (ck/CH/LLN101135 strain, wherein every plumage part vaccine is in deactivation provirus content>=10 for subcutaneous or intramuscular inoculation LX4 type (QX sample) the infectious bronchitis inactivated vaccine of test group
6eID
50) 1 plumage part (0.5ml), the physiological saline of control group subcutaneous vaccination equivalent, observes 14; Cut open inspection, observe whether occur any part of occurring because of vaccinate and systemic reaction or the special pathology of chicken infectious bronchitis as: in chicken tracheae, nasal sinus, have serosity or Catarrhal secretory product, kidney swelling, or occur that typical " piebald kidney " changes.
2.1.2 single dose repeated inoculation test
50 6 Japanese instar chicklings for the proof test of single dose repeated inoculation, every batch of vaccine uses 10, separately establishes 10 of control group chickens, and test group is with subcutaneous or intramuscular inoculation vaccine 1 plumage part, and every chicken inoculated the vaccine of same dose after 14 days with identical route of inoculation.The physiological saline of control group subcutaneous injection equivalent, observes 14; Cut open inspection, whether observation there is any part and systemic reaction or the special pathology of chicken infectious bronchitis that occur because of vaccinate.
2.1.3 heavy dose of inoculation group test
Heavy dose of inoculation test shares 30 6 age in days SPF chick, 2012001 and 2012002 batches of vaccines are each separately establishes 10 of control group chickens with 10, the subcutaneous or intramuscular inoculation vaccine 2 plumage parts (1ml) of test group, the physiological saline of control group injection equivalent, observes 14.Cut open inspection, whether observation there is any part and systemic reaction or the special pathology of chicken infectious bronchitis that occur because of vaccinate.
The proof test of 2.2 kinds of chickens
2.2.1 single dose inoculation test
Single dose inoculation proof test is with 50 AA kind chickens, 10 of the every batch of vaccine uses, and 10 of control group chickens, test group subcutaneous vaccination vaccine 1 plumage part, the physiological saline of control group subcutaneous injection equivalent, observes 30, measures during this time the indexs such as its egg number and hatching rate.
2.1.2 single dose repeated inoculation test
The proof test of single dose repeated inoculation is with 50 AA kind chickens, every batch of vaccine uses 10,10 of control group chickens, the subcutaneous or intramuscular inoculation vaccine 1 plumage part (0.5ml) of test group, every chicken after 14 days with the vaccine of identical route of inoculation repeated inoculation same dose.The physiological saline of control group subcutaneous injection equivalent, observes 30, measures during this time the indexs such as its egg number and hatching rate.
2.1.3 heavy dose of inoculation group test
Heavy dose of inoculation test shares 50 AA kind chickens, and every batch of vaccine uses 10, separately establishes 10 of control group chickens, the subcutaneous or intramuscular inoculation vaccine 1ml of test group, and the physiological saline of control group injection equivalent, observes 30, measures during this time the indexs such as its egg number and hatching rate.
3 experimental results
Under laboratory condition, the safety experiment result of 4 batches of these vaccines (lot number 2012001,2012002,2012003,2012004) shows, 1 age in days and 6 these vaccines of age in days SPF chicken neck subcutaneous injection 0.5ml, inoculate after 14 days, injection site and whole body be without any untoward reaction, proves that chick can use this vaccine safely; The SPF chicken of 6 ages in days, inoculates with different approaches: the injection of the subcutaneous or chest muscle of neck, after every this vaccine of injection 1ml (2 plumage part), to observe 14, injection site absorbs good, and chicken is without systemic adverse reactions, and confirmation different approaches is inoculated all safety; With the vaccinization 2 times after 1 week at interval of 6 age in days SPF chickens, to observe 14, chicken injection site and whole body, without any untoward reaction, illustrate that this vaccine can repeat to use safely; This vaccine is inoculated kind chicken in high term with 1ml/ plumage (2 plumage part), observe 2 months, found that, chicken group is less to the stress reaction of vaccine, egg productivity impact is little, contrast various production performance index no significant differences (P > 0.05) with nonimmune group.
3.1 SPF chick proof tests
From the inoculation test of SPF chick single dose, the test of single dose repeated inoculation, heavy dose of test-results, chicken infectious bronchitis living vaccine is safe and reliable to the SPF chick of 3 ages in days, and physiological activity is normal, cuts open inspection without the special pathology of infectious bronchitis; Proved that this vaccine recommends minimum age in days chicken safe and reliable to vaccine, the searching for food of chicken, drinking-water, daily routines are unaffected.Test group and control group testing data no significant difference.Testing data refers to table 3, table 4 and table 5.
Table 3 inactivated vaccine for avian infectious bronchitis chick single dose inoculation proof test
The proof test of table 4 inactivated vaccine for avian infectious bronchitis chick single dose repeated inoculation
The heavy dose of proof test of table 5 inactivated vaccine for avian infectious bronchitis chick
3.2 kinds of chicken proof tests are from the inoculation test of AA kind chicken single dose, the test of single dose repeated inoculation, heavy dose of test-results; inactivated vaccine for avian infectious bronchitis is safe and reliable to kind of chicken; physiological activity is normal, and the searching for food of chicken, drinking-water, daily routines are unaffected.Test group and control group test egg number and hatching rate no significant difference.Testing data refers to table 6, table 7 and table 8.
Table 6 inactivated vaccine for avian infectious bronchitis kind chicken single dose inoculation proof test
The proof test of table 7 inactivated vaccine for avian infectious bronchitis kind chicken single dose repeated inoculation
The heavy dose of proof test of table 8 inactivated vaccine for avian infectious bronchitis kind chicken
4 experiment conclusion
4.1 inactivated vaccine for avian infectious bronchitis of the present invention (ck/CH/LLN101135 strain) are safe to chick.
4.2 inactivated vaccine for avian infectious bronchitis of the present invention (ck/CH/LLN101135 strain) are safe to kind of the chicken of laying eggs.
The immune duration experiment of embodiment 4 vaccine strain ck/CH/LLN101135 of the present invention strain
1 materials and methods
1.1 vaccine
The inactivated vaccine for avian infectious bronchitis of preparing according to the method described in embodiment 2: lot number is 2012001; Fundamental immunity uses Mass type infectious bronchitis living vaccine H120 purchased from Harbin Wei Ke biotechnology development company.
Chicken is used in 1.2 tests
1 age in days SPF chick.
1.3 attack with strong poison
The strong poison (10 of ck/CH/LLN101135 strain
6.0eID
50/ 0.1ml) be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, its preserving number is: CGMCC No.8790.
1.4 laboratory animal groupings
1 200 of age in days SPF chick, are divided into 10 groups at random, and 20 every group, wherein 10 groups of inactivated vaccine booster immunization groups, 10 groups of fundamental immunity control groups.
1.5 immune
With all 10 groups of chickens of Mass type infectious bronchitis living vaccine H120 immunity, according to the inoculation of vaccine working instructions, 1 plumage part/only; After living vaccine fundamental immunity 30 days with wherein 5 groups of 2012001 batches of inactivated vaccine booster immunizations, remain 5 groups of chick as fundamental immunity control group.The dosage of booster immunization group immune vaccine is plumage part (0.5ml), and control group chicken is 30 days neck subcutaneous vaccination 0.5ml physiological saline after living vaccine fundamental immunity.
Virucidin's dynamic monitoring after 1.6 immunity
1 group of booster immunization chicken and 1 group of fundamental immunity contrast chicken respectively after booster immunization 20 days each group catch and kill 10 chickens, blood sampling separation of serum, measure the NAT of booster immunization chicken and the fundamental immunity control group chicken of identical age in days by virus neutralization tests method, statistics.Catch and kill and will remain chicken attack ck/CH/LLN101135 strain poison by force the same day, attack toxic agent amount and be no less than 10
5.0eID
50/ only, attack poison and within latter 5 days, catch and kill these two groups all residue chickens, collect respectively serum, to measure and tire for the neutralization of ck/CH/LLN101135 strain virus, statistics gathers every chicken kidney and tracheae simultaneously, carries out viral separation detection, statistics virus separation rate again.
The gentle pipe virus of kidney separation rate again after remaining 4 groups of booster immunization chickens and 4 groups of fundamental immunities contrast chickens and measuring serum NATs in 60,90,120,150 days after booster immunization respectively and attack poison with program as stated above, the immune duration providing after the immunity of infectious bronchitis inactivated vaccine is provided.
2 experimental results
2.1 antibody extended periods
2012001 batches of vaccines with the dosage immunized chicks of 1 plumage part after 20,60,90,120 days, the inactivated vaccine booster immunization chicken serum NAT all high 2 times of above chicken quantity of highest serum NAT of the fundamental immunity contrast chicken of more identical age in days only reaches 6-9, and the serum NAT of inactivated vaccine booster immunization chicken all contrasts the average serum NAT of chicken higher than fundamental immunity.Simultaneously, 10 chicken serum NATs of attacking poison after inactivated vaccine booster immunization are attacked the high 2 times of above chicken quantity of malicious fundamental immunity contrast chicken serum NAT compared with 10 and are only reached 6-9, and the serum NAT of inactivated vaccine booster immunization chicken is all higher than the average serum NAT of fundamental immunity contrast chicken.Attack malicious chicken target organ virus separation detection result and show, booster immunization chicken tracheae separates positive rate than the low 20-30% of fundamental immunity control group (2-3/10) with virus in kidney.And after deactivation vaccine booster immunization chick 150 days, the high 2 times of above chicken quantity of highest serum NAT of the fundamental immunity contrast chicken of the more identical age in days of inactivated vaccine booster immunization chicken serum NAT only reach 4, and the serum NAT of inactivated vaccine booster immunization chicken all contrasts the average serum NAT of chicken higher than fundamental immunity.Simultaneously, 10 chicken serum NATs of attacking poison after inactivated vaccine booster immunization are attacked the high 2 times of above chicken quantity of malicious fundamental immunity contrast chicken serum NAT compared with 10 and are reached 5, and the serum NAT of inactivated vaccine booster immunization chicken is all higher than the average serum NAT of fundamental immunity contrast chicken.Attack malicious chicken target organ virus separation detection result and show, booster immunization chicken tracheae and in kidney virus to separate positive rate lower by 10% (1/10) than fundamental immunity control group.The results are shown in Table 9-13; from the visible infectious bronchitis inactivated vaccine of result booster immunization, higher than control group, more than 2 times chicken quantity does not reach 5 to 150 days NATs; after vaccine immunity is described, NAT declines; immunoprotection Efficiency Decreasing; Given this, the immune duration of infectious bronchitis inactivated vaccine is decided to be to 4 months.
Latter 20 days booster immunization groups of table 9 immunity and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
Latter 60 days booster immunization groups of table 10 immunity and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
Latter 90 days booster immunization groups of table 11 immunity and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
Latter 120 days booster immunization groups of table 12 immunity and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
Latter 150 days booster immunization groups of table 13 immunity and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
3 experiment conclusion
The vaccine immunity extended period of the present invention is 4 months.
The immune efficacy contrast experiment of embodiment 5 LX4 type of the present invention (QX sample) infectious bronchitis inactivated vaccine strain ck/CH/LLN101135 strain and Mass type infectious bronchitis inactivated vaccine strain M41 strain
1, materials and methods
1.1 fundamental immunity vaccines
Mass type infectious bronchitis living vaccine (H120 strain) and infectious bronchitis inactivated vaccine (M41 strain) are all purchased from Harbin Wei Ke biotechnology development company, vaccine preservation condition and validity period :-15 DEG C of following preservations, validity period is 12 months.Press label mark plumage part with drawing vaccine after sterile saline dilution, every chicken eye droppings collunarium is inoculated 1 plumage part.
1.2 the preparation of LX4 type (QX sample) infectious bronchitis inactivated vaccine
According to embodiment 2 method preparations.
The effect evaluation of 1.3 infectious bronchitis inactivated vaccines
40 1 age in days SPF chickens are divided into 4 groups (20 every group) at random, in negative pressure isolator, raise and whole immune Mass type infectious bronchitis living vaccines (H120 strain), and 1 plumage part/only.Wherein the 1st group of chick exempted from latter 20 days subcutaneous inoculation inoculation LX4 type (QX sample) infectious bronchitis inactivated vaccines in H120 head (ck/CH/LLN101135 strain, wherein every plumage part vaccine is>=10 at deactivation provirus content
6eID
50), every subcutaneous vaccination 0.5ml.Exempt from latter 20 days subcutaneous inoculations inoculation Mass type infectious bronchitis inactivated vaccine (M41 strain) in H120 head for the 2nd group, every subcutaneous vaccination 0.5ml, the other the 3rd and 4 groups of chick respectively as H120 living vaccine fundamental immunity control group.Booster immunization group is respectively got 10 chickens blood samplings separation of serum in two groups of immunity rear 20 days, and application serum neutralization test method detects the serum NAT of every chicken.Simultaneously by the 1st and 3 groups of residue chick with strong malicious ck/CH/LLN101135 or carry out eye droppings and collunarium attack, attack toxic agent amount for every and be no less than 10
5.0eID
50attack poison and within latter 5 days, catch and kill booster immunization group and fundamental immunity control group chicken, blood sampling separation of serum respectively, gather kidney, the tracheae of every chicken simultaneously, the serum gathering is measured respectively for the neutralization of ck/CH/LLN101135 strain and tired, measure the IBV target organ tracheae and the kidney that gather simultaneously, carry out virus isolated rate mensuration.Booster immunization group serum antibody titer and fundamental immunity contrast the difference between chicken, the effect of evaluation infectious bronchitis inactivated vaccine before attacking poison by contrast and after attacking poison.Corresponding, by the 2nd and 4 groups of residue chick with strong malicious M41 or carry out eye droppings and collunarium attack after with synchronous method measure for the neutralization of ck/CH/LLN101135 strain tire attack poison by contrast before and attack the rear booster immunization group serum antibody titer of poison and fundamental immunity contrasts the difference between chicken, the immune efficacy of evaluation Mass type infectious bronchitis inactivated vaccine to LX4 type (QX sample) strain.
2 results
2.1 LX4 type (QX sample) infectious bronchitis inactivated vaccine immune efficacy evaluation result
After Mass type living vaccine head exempts from, after LX4 type (QX sample) infectious bronchitis inactivated vaccine booster immunization, 10 chicken serum NATs of 20 days reach 9 compared with the high more than 2 times chicken quantity of highest serum NAT of 10 fundamental immunity contrast chickens, and the serum NAT of inactivated vaccine booster immunization chicken all contrasts the average serum NAT of chicken higher than fundamental immunity.Simultaneously, 10 chicken serum NATs of attacking poison after inactivated vaccine booster immunization are attacked the high 2 times of above chicken quantity of malicious fundamental immunity contrast chicken serum NAT compared with 10 and are reached 9, and the serum NAT of inactivated vaccine booster immunization chicken is all higher than the average serum NAT of fundamental immunity contrast chicken.Attack malicious chicken target organ virus separation detection result and show, booster immunization chicken tracheae separates positive rate than the low 20-30% of fundamental immunity control group (2-3/10) with virus in kidney.The results are shown in Table 14.
2.3 Mass type infectious bronchitis inactivated vaccine (M41 strain) immune efficacy evaluation result
After Mass type living vaccine head exempts from, after Mass type (M41 strain) infectious bronchitis inactivated vaccine booster immunization, 10 chicken serum NATs of 20 days are compared with only 1 of the high more than 2 times chicken quantity of highest serum NAT of 10 fundamental immunities contrast chickens, and the serum NAT of inactivated vaccine booster immunization chicken does not contrast the average serum NAT of chicken higher than fundamental immunity.After Mass type (M41 strain) inactivated vaccine booster immunization, attack 10 chicken serum NATs of poison and attack only 3 of the more than high 2 times chicken quantity of malicious fundamental immunity contrast chicken serum NAT compared with 10.Attack malicious chicken target organ virus separation detection result and show, booster immunization chicken tracheae and in kidney virus separate positive rate and fundamental immunity control group all at 80-90%.The results are shown in Table 15.
Table 14 LX4 type (QX sample) infectious bronchitis vaccine booster immunization group and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
Table 15 Masss type infectious bronchitis vaccine booster immunization group and the neutralization of fundamental immunity control group serum antibody are tired and target organ virus isolated rate
3. conclusion (of pressure testing)
LX4 type (QX sample) infectious bronchitis inactivated vaccine ck/CH/LLN101135 strain booster immunization can the neutralizing antibody level of effective stimulus body lift body acupuncture to LX4 type (QX sample) infectious bronchitis virus, illustrate that ck/CH/LLN101135 strain infectious bronchitis inactivated vaccine of the present invention more current only commercial goods infectious bronchitis inactivated vaccine (Mass type) immune effect is better, after immunity, excite the neutralizing antibody producing for China's Major Epidemic strain, promote chicken group health level.
Claims (3)
1. avian infectious bronchitis virus vaccine strain, called after ck/CH/LLN101135, is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and its preserving number is: CGMCC No.8790.
2. avian infectious bronchitis virus vaccine strain claimed in claim 1 is in the application of preparing in inactivated vaccine for avian infectious bronchitis.
3. prevent and treat a vaccine composition for chicken infectious bronchitis, it is characterized in that being formed by avian infectious bronchitis virus vaccine strain claimed in claim 1 and pharmaceutically acceptable immunological adjuvant after deactivation.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105624122A (en) * | 2016-02-01 | 2016-06-01 | 浙江大学 | Avian infectious bronchitis virus natural mutant |
| CN105802918A (en) * | 2014-12-29 | 2016-07-27 | 普莱柯生物工程股份有限公司 | Avian nephropathogenic infectious bronchitis virus strain as well as vaccine composition, preparation method and application thereof |
| CN109239342A (en) * | 2018-10-19 | 2019-01-18 | 中国农业大学 | QX type IBV hemagglutination-inhibition test antigen and the preparation method and application thereof |
| CN109628412A (en) * | 2018-12-27 | 2019-04-16 | 山东省农业科学院家禽研究所（山东省无特定病原鸡研究中心） | One plant of avian infectious bronchitis virus strain and its application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001009290A2 (en) * | 1999-07-30 | 2001-02-08 | Laboratorio Avi-Mex, S.A. De C.V. | Avian infectious bronchitis virus serotype |
| TW200710224A (en) * | 2005-09-02 | 2007-03-16 | Univ Nat Taiwan | Attenuated avian infectious bronchitis virus vaccines |
| CN101514334A (en) * | 2009-03-23 | 2009-08-26 | 中国农业科学院哈尔滨兽医研究所 | Chicken infectivity bronchitis virus attenuated vaccine strain and application thereof |
| CN102220287A (en) * | 2011-04-29 | 2011-10-19 | 中国农业科学院哈尔滨兽医研究所 | Avian infectious bronchitis cold adaptation attenuated vaccine strain and application thereof |
-
2014
- 2014-07-10 CN CN201410327912.XA patent/CN104130981A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001009290A2 (en) * | 1999-07-30 | 2001-02-08 | Laboratorio Avi-Mex, S.A. De C.V. | Avian infectious bronchitis virus serotype |
| TW200710224A (en) * | 2005-09-02 | 2007-03-16 | Univ Nat Taiwan | Attenuated avian infectious bronchitis virus vaccines |
| CN101514334A (en) * | 2009-03-23 | 2009-08-26 | 中国农业科学院哈尔滨兽医研究所 | Chicken infectivity bronchitis virus attenuated vaccine strain and application thereof |
| CN102220287A (en) * | 2011-04-29 | 2011-10-19 | 中国农业科学院哈尔滨兽医研究所 | Avian infectious bronchitis cold adaptation attenuated vaccine strain and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| CHUYANG SUN ET AL.: "Phylogenetic analysis of infectious bronchitis coronaviruses newly isolated in China, and pathogenicity and evaluation of protection induced by Massachusetts serotype H120 vaccine against QX-like strains", 《AVIAN PATHOLOGY》 * |
| GENBANK: "GenBank：HM194640.1", 《GENBANK》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105802918A (en) * | 2014-12-29 | 2016-07-27 | 普莱柯生物工程股份有限公司 | Avian nephropathogenic infectious bronchitis virus strain as well as vaccine composition, preparation method and application thereof |
| CN105802918B (en) * | 2014-12-29 | 2019-11-19 | 普莱柯生物工程股份有限公司 | Chicken's infectious bronchitis nephritis strain and its vaccine composition, preparation method and application |
| CN105624122A (en) * | 2016-02-01 | 2016-06-01 | 浙江大学 | Avian infectious bronchitis virus natural mutant |
| CN109239342A (en) * | 2018-10-19 | 2019-01-18 | 中国农业大学 | QX type IBV hemagglutination-inhibition test antigen and the preparation method and application thereof |
| CN109239342B (en) * | 2018-10-19 | 2020-06-09 | 中国农业大学 | QX type IBV hemagglutination inhibition test antigen and preparation method and application thereof |
| CN109628412A (en) * | 2018-12-27 | 2019-04-16 | 山东省农业科学院家禽研究所（山东省无特定病原鸡研究中心） | One plant of avian infectious bronchitis virus strain and its application |
| CN109628412B (en) * | 2018-12-27 | 2021-11-02 | 山东省农业科学院家禽研究所（山东省无特定病原鸡研究中心） | Infectious bronchitis virus strain and application thereof |
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