CN102086446A - Strain of Adenoviridae and application thereof - Google Patents
Strain of Adenoviridae and application thereof Download PDFInfo
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Abstract
The invention discloses a strain of Adenoviridae, CGMCC NO:3527. The strain is a novel separated strain of Adenoviridae and same in serotype with an international standard strain AV127. The strain has good virus immunogenicity and can resist attach from separated strains and standard strains of Adenoviridae in multiple places. An immunize egg laying chicken flock of an oil emulsion of Adenoviridae can protect the laying rate of the chicken flock and is an ideal seedling candidate strain. The strain can be used for preparing specific EDS76 virus antigen and positive serum for rapid verification of virus and investigation in epidemiology.
Description
Technical field
The invention belongs to the microbial virus field, relate to the new seed culture of viruses of virus, be specifically related to a stud bird egg drop syndrome virus stain and an application thereof.
Background technology
The Luo Man father and mother that the Henan chicken house is raised are for kind of a chicken, and in 31 ages in week, this chicken group does not inoculate the egg drop syndrome inactivated vaccine.On February 12nd, 2002, the consumption of chicken group feeding material is normal, and egg productivity descends significantly.Premorbid day, laying rate was 86.7%, and morbidity back egg productivity linearly descends, and drops to lower-most point on 9th, is 45.6%.After this, slowly ging up appears in egg productivity, the 30th everyday laying rate reach 52.7%, still can not return to premorbid level.Between period of disease, an average day laying rate is 50.1%, has descended 36.5% than premorbid, observes the phenomenon of producing soft shelled egg, shell egg and thick shell egg simultaneously, and these lopsided eggs account for 11.5% of total egg number.
The how no abnormal performance of ill chicken group mental status, only the minority chicken is suffered from diarrhoea, and is total to 6 of dead chickens between period of disease, and mortality ratio is 0.6 ‰.Cut open the chicken organ organ of dying of illness extremely and do not have obvious naked eyes variation, the blood of coring, liver, the also negative result of spleen bacteriology checking.Gather 19 parts in the back 12 days chicken serum sample of morbidity, detect EDS
76Hemagglutination inhibition antibody, positive rate is 100%, tiring is 2
5～2
8
Summary of the invention
The purpose of this invention is to provide a stud bird egg drop syndrome virus stain, classification name: fowl egg drop syndrome virus, Latin formal name used at school: Adenoviridae, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, depositary institution is called for short: CGMCC, depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, preservation date: on December 17th, 2009, deposit number: 3527.
Technical scheme of the present invention is: a stud bird egg drop syndrome virus stain, fowl egg drop syndrome virus (Adenoviridae), CGMCC NO:3527.
Described fowl egg drop syndrome virus stain has following characteristics:
(1) described strain shows the virus particle that is sexangle, sphere, no cyst membrane, diameter 75～80nm under Electronic Speculum, meets the morphological specificity of adenovirion;
(2) described strain is insensitive to chloroform and ether, and at pH being has resistibility under 3.0～10.0 the condition, under 37～70 ℃ condition resistibility is arranged;
(3) nucleic acid type of described strain is a dna virus;
(4) described strain only with EDS76 antiserum(antisera) generation specific reaction, and be 0 with the HI reaction of H5, H7, H9 and ND positive serum;
(5) described strain has only EDS in duck embryo neutralization test
76Antiserum(antisera) can suppress the propagation of described strain virus at duck embryo allantoic cavity, and other positive serum all can not suppress the propagation of described strain virus;
(6) the relationship value R of described strain and international standard strain AV127 is 95%, belongs to same serotype;
(7) use the egg drop syndrome oil emulsion adjuvant emulsion of described strain preparation can stimulate chicken to produce EDS76 antibody.
The application of fowl egg drop syndrome virus stain of the present invention in the production seed culture of viruses of preparation vaccine.
Fowl egg drop syndrome virus stain of the present invention is at preparation fowl egg drop syndrome viral diagnosis EDS
76Application in the antigen reagent.
The application of fowl egg drop syndrome virus stain of the present invention in preparation fowl egg drop syndrome viral diagnosis positive serum reagent.
Fowl egg drop syndrome virus stain of the present invention is at preparation fowl egg drop syndrome viral therapy EDS
76Application in the antiserum(antisera) reagent.
The invention has the beneficial effects as follows: the present invention is the new egg drop syndrome virus isolated strain of a strain, and is identical with the serotype of international standard strain AV127 strain; Virus immunity originality is good, can resist the attack of egg drop syndrome a plurality of Local Isolates of virus and type strain; Egg drop syndrome virus oil-emulsion immunity laying hen group of the present invention provides effective protection can for chicken group's laying rate, is the more satisfactory seedling candidate strain of a strain; Can be used for preparing specific EDS
76Virus antigen and positive serum are used for the Rapid identification and the EPDML investigation of virus.
Description of drawings
Fig. 1 is the target gene PCR amplification,
Wherein, 1.1kb DNA ladder marker (8000,7000,6000,5000,4000,3000,2000,1500,1000,517,396and 230bp); 2. target gene PCR product.
Fowl egg drop syndrome virus stain of the present invention, preservation date: on December 17th, 2009, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, depositary institution is called for short: CGMCC, deposit number: 3527.
Embodiment
Embodiment
In February, 2002, certain chicken house laying hen group egg drop reduction of Henan is gathered pathological material of disease, is separated to a strain egg drop syndrome virus stain after duck embryonic breeding kind.Chicken group 190 ages in days begin morbidity, descending and produce lopsided eggs such as light egg, soft shelled egg, shell egg with egg productivity is principal character, others such as morbidity chickens' extract god, appetite are normal substantially, and above-mentioned symptom of facing the chicken egg drop syndrome after examining performance and falling ill in other areas of domestic report in recent years is roughly similar.
When described strain separated for the first time, duck embryo allantoic liquid blood clotting titre was 0, but reached 2 during to the 3rd generation
9, through continuous passage to the during 5 generations HA tire stable and can reach 2
15～17, and the red corpuscle of its aggegation chicken, and the mammiferous red corpuscle of not aggegation.
The effect of the aggegation chicken red blood cell of described strain is not suppressed by NDV and AIV antiserum(antisera), but can be by EDS
76Positive serum suppresses.
Described strain is after infecting laying hen, and the test chicken egg productivity descends and unusual egg increases.
Electron microscopic observation can be seen typical adenovirion.
The physicochemical property test shows that described strain is insensitive to fatsolvent, and soda acid and heat are all had certain resistibility.
Cytopathy can be bred and occur to virus on DEF; The nucleic acid type of virus is a dna virus; PCR result shows that also described strain virus is a strain egg drop syndrome virus.
Described strain has good specificity and immunogenicity.Virus liquid and equivalent EDS
76Positive serum mixed 37 ℃ of effects after 1 hour, the inoculation DEF, and acellular pathology occurs, and it is 0 that HA tires.And ND, AIV positive serum can not suppress the propagation of virus, illustrate that described strain specific is strong.
The result of neutralization test shows that the serological relation of the serotype of described strain and international standard strain AV127 is close, belongs to same serotype.Dye chick and oil-emulsion immunity test chicken with viral liquid inductance, test chicken has all produced specificity EDS
76HI antibody, the chicken egg drop syndrome virus stain that is separated to from serology angle proof has good immunogenicity.
The application of fowl egg drop syndrome virus stain of the present invention in the production seed culture of viruses of preparation vaccine.
Fowl egg drop syndrome virus stain of the present invention is at preparation fowl egg drop syndrome viral diagnosis EDS
76Application in the antigen reagent.
The application of fowl egg drop syndrome virus stain of the present invention in preparation fowl egg drop syndrome viral diagnosis positive serum reagent.
Fowl egg drop syndrome virus stain of the present invention is at preparation fowl egg drop syndrome viral therapy EDS
76Application in the antiserum(antisera) reagent.
Among the present invention, strain is named the strain into Chicken/China/Henan/02/2002, is called for short the HE02 strain.
(1) epidemiology survey: investigation chicken group's age of onset, day laying rate, eggshell quality, situations such as clinical manifestation and death.The Luo Man father and mother that the Henan chicken house is raised are for kind of a chicken, and in 31 ages in week, this chicken group does not inoculate the egg drop syndrome inactivated vaccine.On February 12nd, 2002, the consumption of chicken group feeding material is normal, and egg productivity descends significantly.Premorbid day, laying rate was 86.7%, and morbidity back egg productivity linearly descends, and drops to lower-most point on 9th, is 45.6%.After this, slowly ging up appears in egg productivity, the 30th everyday laying rate reach 52.7%, still can not return to premorbid level.Between period of disease, an average day laying rate is 50.1%, has descended 36.5% than premorbid, observes the phenomenon of producing soft shelled egg, shell egg and thick shell egg simultaneously, and these lopsided eggs account for 11.5% of total egg number.
The how no abnormal performance of ill chicken group mental status, only the minority chicken is suffered from diarrhoea, and is total to 6 of dead chickens between period of disease, and mortality ratio is 0.6 ‰.Cut open the chicken organ organ of dying of illness extremely and do not have obvious naked eyes variation, the blood of coring, liver, the also negative result of spleen bacteriology checking.Gather 19 parts in the back 12 days chicken serum sample of morbidity, detect EDS
76Hemagglutination inhibition antibody, positive rate is 100%, tiring is 2
5～2
8
(2) virus is separated: get die of illness chicken salpingo and uterus and add 10 times of physiological saline respectively and grind homogenate; Other gets soft shelled egg egg white and adds an amount of physiological saline and prepare the egg white diluent.The chloroform that in above-mentioned 3 parts of pathological material of diseases, adds equivalent, jolting 30 minutes, then, and with 3500rpm centrifugal 10 minutes, get supernatant liquor through two anti-processing, 4 ℃ are spent the night.Second day, pathological material of disease is made sterility test, get aseptic material as viral separating sample, put one 30 ℃ of refrigerators and freeze standby.
3 increments are originally inoculated the duck embryo and the 10 age in days SPF chicken embryos of 10～11 ages in days respectively, and the duck embryo is from EDS
76The duck group of hemagglutination inhibition antibody feminine gender, through the inoculation of allantoic cavity approach, every embryo 0.2ml was hatched 96～120 hours continuously in 37 ℃, and per 6～8 hours photograph eggs are once in time chosen dead germ; Discard the dead germ before 48 hours, no matter all embryos are taken out in dead or survival after 120 hours, air chamber is upwards upright, puts 2～8 ℃ of refrigerator coolings 8～16 hours, the results blastochyle, observe the mortality ratio of each generation duck embryo and chicken embryo, due to pathology and measure the hemagglutinative titer of allantoic fluid.And carry out blind passage, in per 1 generation, all established 2 pieces of duck embryos of not inoculating in contrast.
The result shows, will die of illness chicken salpingo and uterus two this inoculation of increment duck embryos and 3 generations of blind passage, the duck embryo and produce pathology of all can not causing death, and the allantoic fluid of measuring results does not also have hemagglutinative titer.
Soft shelled egg egg white sample is inoculated the duck embryo and carried out blind passage, and during to the 3rd generation, 3/10 duck embryo incidence is observed a small amount of petechial hemorrhage, occurs 2 in the allantoic fluid of results
4～7The blood clotting valency.When being passaged to the 4th generation and the 5th generation, at back 118 hours and 115 hours 1 piece of duck embryos that causes death of inoculation, mortality ratio is 20% (2/10) respectively; Dead embryo is remarkable with survival embryo pathology, the fetal development retardance, and idiosome incidence, back, thigh, wing and belly are hemorrhage obviously, and severe patient is diffuse hemorrhage; The blood clotting valency of results allantoic fluid is from 2
7～2
11Rise to 2
13～2
17In 5 processes that go down to posterity, the healthy survival of contrast duck embryo, the allantoic fluid undetermined goes out the blood clotting valency.Virus is separated the chicken embryo of acquisition and the HA and the HI test-results of duck embryo allantoic liquid sees Table 1,2.
Table 1 pathological material of disease isolate uses the test-results of chicken red blood cell
Table 2 pathological material of disease isolate uses the test-results of rabbit erythrocyte
(3) artificial challenge's test of falling ill: collect 56 pieces of thick shell eggs, shell egg and soft shelled eggs from ill chicken group, smash the back mix fodder to pieces and directly feed EDS
76Healthy 30 of the commodity eggs of Luo Man (190 age in days) of negative antibody, continuous 3 days, are raised in the shield retaining at every day 1 time.Feed back the 0th, 8,15 and 27 days from every test chicken blood sampling separation of serum, measure EDS respectively
76And newcastle disease (ND) hemagglutination inhibition antibody tires, simultaneously the mental status, the appetite of viewing test chicken, drink water and lay eggs situation.Other establishes with 30 chickens of age in days and contrasts as non-infection.
The result shows that after the content artificial challenge with thick shell egg, shell egg and soft shelled egg, the mental status of test chicken, appetite and drinking-water do not have any ANOMALOUS VARIATIONS.The 8th, all serology has taken place infected chicken changeed sun, EDS
76Hemagglutination inhibition antibody is determined as 2
6～2
9The 15th, serum antibody titer rose to 2
9～2
10, this high antibody horizontal lasted till the 30th always.In this trial period, obvious variation does not take place in chicken group's ND hemagglutination inhibition antibody.Behind the artificial challenge the 16th day, the test group hen gave birth to the 1st piece of soft shelled egg, after this collects 43 pieces of thick shell eggs, 23 pieces of lopsided eggs and 32 pieces of soft shelled eggs again, and these unusual eggs account for 27.2% (98/360) of total egg number.
After artificial challenge's off-test, cut open the hen that produces soft shelled egg extremely, gather the egg white material of uterine tube, uterus and soft shelled egg respectively, inoculate EDS once more after handling by preceding method
76Negative duck embryo is promptly measured higher hemagglutinative titer in the allantoic fluid of the 1st pickup kind duck embryo.The hemagglutinative titer of 3 kinds of material duck embryo isolates reaches 2 respectively
8, 2
6With 2
9
(4) virus PCR is identified: with reference to the conserved regions of the pVIII protein gene of the egg drop syndrome virus X99782 that delivers on the GenBank, the synthetic following primer of design, upstream primer P1:5 '-TCA
GAA TTCTGT TGG AGC TGA TTA GGAAGG-3 ', the line sequence is an EcoR I restriction enzyme site; Downstream primer P2:5 '-AGT
AAG CTTGGT GGAAAT TCA TGT ACAAAC-3 ' is scribed ss the HindIII restriction enzyme site.This strides the width of cloth to the theory between primer is 1348bp.
Extract virus genomic DNA.
Pcr amplification.Viral nucleic acid with extraction is a template, reaction system 25 μ l, and reaction system is: 10 * Buffer2.5 μ l, dNTP (2.5mol/L) 2 μ l, Ex TaqDNA polysaccharase (Promega) 0.12 μ l (5U μ l
-1), each 1 μ l of upstream and downstream primer, template 2 μ l, replenishing volume with distilled water is 25 μ l, reaction conditions is as follows: 98 ℃, 5 minutes, 72 ℃, 1 minute, 1 circulation added ExTaq polysaccharase (TaKaRa company); 94 ℃ of pre-sex change 5 minutes, 1 minute, 62 ℃ annealing of 94 ℃ of sex change were extended totally 35 circulations 1.5 minutes for 1 minute, 72 ℃; Last 72 ℃ are extended end reaction in 10 minutes, get product 5 μ l and detect amplification through 1.5% agarose gel electrophoresis.
Above-mentioned PCR product electrophoresis on 1.5% sepharose, cutting purpose fragment under ultraviolet lamp reclaims test kit with glue then and carries out DNA recovery and purifying, and operation is undertaken by explanation, and it is standby that the recovery product is put-20 ℃ of preservations.
The clone of target gene PCR product and sequencing.Purpose fragment that will be through reclaiming purifying and T carrier (pGEM-T easy) by the pGEM-T easy Vector of Promega company illustrate be connected after, be built into recombinant plasmid pEDS, with its transformed competence colibacillus cell JM109, coating LB flat board is at last by blue hickie preliminary screening positive colony.Get the positive bacterium colony that filters out and shake bacterium, carry out the extraction of plasmid, the recombinant plasmid that extracts is carried out PCR, EcoR I, Hind III single endonuclease digestion and double digestion identify 1.5% gel agarose electrophoretic analysis.Send precious biotechnology (Dalian) company limited to carry out sequencing analysis the bacterial strain that is accredited as positive colony.Utilize the BLAST among the GenBank that sequencing result is carried out the sequence alignment analysis subsequently.
The result shows, as shown in Figure 1, is template with the DNA of this strain isolated, and amplification pVIII protein gene part fragment after the electrophoretic analysis, amplifies the purpose band of 1348bp, conforms to expected result; Pass through sequencing, record the long 1348bp of being of goal gene nucleotide sequence, as the present invention relates to shown in the nucleotides sequence tabulation, conform to fully with the corresponding gene fragment of the gene order of having delivered (X99782) among the GenBank, verified that further the cause of disease that is separated to is an egg drop syndrome virus.
(5) blood clotting (HA) and mutual hemagglutination-inhibition test (HI) test: use red corpuscle of chicken and the allantoic fluid of red corpuscle of rabbit, adopt micromethod on 96 hole V-type Sptting plates, to carry out the HA test according to a conventional method to gathering in the crops; Anti-EDS is used in the HI test
76AV127 standard positive serum and duck embryo allantoic liquid, anti-NDV standard positive serum and duck embryo allantoic liquid carry out on 96 hole V-type Sptting plates according to " Chinese veterinary drug allusion quotation " method.
(6) electron microscopic examination: the viral liquid that will gather in the crops was through 6000r/ minute, and 30 minutes, 15000r/ minute centrifugal 20 minutes, behind the 35000r/ minute differential centrifugation, get precipitation and use sessile drop method negative staining (2%PTA, pH value 6.7) 3 minutes with pH value 7.0PBS liquid washing back, the back electron microscopic observation dries in the shade.
The electron microscopic examination result shows, can see the virus particle that is hexangle type, sphere, no cyst membrane, the about 75～80nm of diameter under Electronic Speculum, meets the morphological specificity of adenovirion substantially.
(7) physicochemical property of virus is measured:
A, chloroform sensitivity test.Virus and chloroform by 4: 1 mixings, are put 37 ℃ of effects 30 minutes, centrifugal 30 minutes then through 4000rpm, get supernatant liquor inoculation DEF, the 0.2ml/ hole, connect aseptic PBS liquid simultaneously and the viral liquid handled without chloroform in contrast.Through 37 ℃ of 5%CO
2Cultivated 96～120 hours, aseptic harvested cell nutrient solution is measured the HA valency.
To ether sensitivity test.With adding the ether vibration 10 minutes that final concentration is 20% (V/V) in the viral liquid, place 4 ℃ to spend the night, ether is removed in volatilization, the inoculation DEF, the 0.2ml/ hole is provided with the contrast of unprocessed viral liquid and aseptic PBS liquid, simultaneously through 37 ℃ of 5%CO
2Cultivated 96～120 hours, aseptic harvested cell nutrient solution is measured the HA valency.
Behind chloroform or ether viral liquid inoculation DEF that handle and without chloroform or ether processing, all can cause the DEF pathology, cell circle before this contracts, and death comes off then, pulls into netted; Its blood clotting valency does not have considerable change.Control group (PBS liquid) is then normal, and the HA valency is zero, illustrates that this virus is insensitive to chloroform and ether.
B, heat stability test.After viral liquid carried out 56 ℃ 3 hours, 60 ℃ 30 minutes, 70 ℃ water-bath effects in 30 minutes respectively and handle, the inoculation DEF, the 0.2ml/ hole is provided with the contrast of unprocessed viral liquid and aseptic PBS liquid, simultaneously through 37 ℃ of 5%CO
2Cultivated 96～120 hours, aseptic harvested cell nutrient solution is measured the HA valency.56 ℃ of water-bath effects in 3 hours of HE02 strain virus liquid can be survived after handling, and cytopathy, HA occur and tire and change not quite, and 60 ℃ 30 minutes, 70 ℃ can make inactivation of virus in 30 minutes.
C, acid resistance test.Get viral liquid 0.1mol/L HCl solution adjust pH to 3.0,4.0,5.0 through 37 ℃ of effects 1 hour, use 7.5%NaHCO then
3Adjust pH to 7.0～7.2, the inoculation DEF, the 0.2ml/ hole is provided with the contrast of unprocessed viral liquid and aseptic PBS liquid, simultaneously through 37 ℃ of 5%CO
2Cultivated 96～120 hours, aseptic harvested cell nutrient solution is measured the HA valency.
Alkali resistance test.Get viral liquid and the pH value is transferred to 8.0 with 0.1mol/L NaOH solution, 9.0,10.0,37 ℃ act on 1 hour, use 0.1mol/L HCl solution adjust pH to 7.0～7.2 then, inoculation DEF, 0.2ml/ hole, the contrast of unprocessed viral liquid and aseptic PBS liquid is set simultaneously, through 37 ℃ of 5%CO
2Cultivated 96～120 hours, aseptic harvested cell nutrient solution is measured the HA valency.
After 1 hour adjust pH to 7.0～7.2 through 37 ℃ of effects through pH 3.0,4.0,5.0,8.0,9.0,10.0 respectively for virus liquid, behind the inoculation DEF, with do not handle viral liquid phase relatively, the cytopathy situation is basic identical, all can cause the DEF pathology, cell circle before this contracts, and death comes off then, pulls into netted; Its HA basic no change of tiring illustrates that this virus particle all has certain resistibility to bronsted lowry acids and bases bronsted lowry.
(8) immunogenicity of strain
With the HE02 strain virus with 2 * 10
4TCID
50Inoculation SPF chicken in 18 age in week, EDS after 9 days
76HI antibody 〉=2
7.5Preparation oil-emulsion immunity SPF chicken in 4 age in week is 10 after the deactivation of virus liquid, and 3～5 weeks of immunity back are gathered serum, measure EDS
76HI tires 〉=and 2
9The results are shown in Table 3.
The infectivity of table 3HE02 strain virus and immunogenicity test
(9) antigen-specific:
With viral liquid respectively with equivalent AI, ND, EDS
76Positive serum mixed 37 ℃ of effects after 1 hour, the inoculation DEF, and observation of cell pathology, mensuration HA tire.EDS
76Positive serum can suppress the propagation of virus, and it is 0 that acellular pathology, HA tire; And ND, AI positive serum can not suppress the propagation of virus.The results are shown in Table 4.
The specificity of table 4HE02 strain virus
(10) neutralization test: the HE02 strain positive serum of AV127 positive serum and preparation put in 56 ℃ of water-baths handled 30 minutes, with keeping 10 times of dilutions of going forward one by one of liquid do, respectively with 100TCID
50Above-mentioned two strain balanced mix, hatched 1 hour for 37 ℃, add the DEF Tissue Culture Plate then successively, each extent of dilution adds 6 holes, cultivate 72 hours observation of cell pathologies, the hemagglutinative titer of 96 hours mensuration cell conditioned medium liquid calculates the neutralization of serum and tires by the Reed-Muench method, and compares the serology dependency between two strains.
Test-results shows, the cross neutralization reaction can take place for HE02 strain and AV127 strain, with a serum to different EDS
76The neutralization of strain is tired and is existed difference.The ratio that neutralization is tired according to allos serum between two strains and homology serum, (r1=Ab/Aa, r2=Ba/Bb), by the formula R=100 (r1r2) 1/2 that proposes in the foreign literature, and R value＞80% o'clock, serotype is identical to draw relation value r1 and r2.Calculate the relationship value R=95% of two strains in view of the above,, show from isolating HE02 strain strain and international standard poison AV127 serological relation closely, belong to same serotype greater than 80%.The results are shown in Table 5.
Table 5HE02 strain strain neutralization test result
＜110〉Agricultural University Of He'nan
＜120〉a stud bird egg drop syndrome virus stain and an application thereof
<160>1
<170>PatentIn?Version?3.2
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<211>1348
<212>DNA
＜213〉fowl egg drop syndrome virus (Adenoviridae)
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gcagggtatg?atttgcatga?tgggcgccag?tataggaaaa?taacaagaga?tgcgttgcca 780
tttcctcata?attggcaggt?caaagatggt?tcgcggtggg?taaatctggg?tgggaaagga 840
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Claims (6)
1. a stud bird egg drop syndrome virus stain is characterized in that: fowl egg drop syndrome virus (Adenoviridae), CGMCC NO:3527.
2. fowl egg drop syndrome virus stain according to claim 1, it is characterized in that: it has following characteristics:
(1) described strain shows the virus particle that is sexangle, sphere, no cyst membrane, diameter 75～80nm under Electronic Speculum, meets the morphological specificity of adenovirion;
(2) described strain is insensitive to chloroform and ether, and at pH being has resistibility under 3.0～10.0 the condition, under 37～70 ℃ condition resistibility is arranged;
(3) nucleic acid type of described strain is a dna virus;
(4) described strain only with EDS76 antiserum(antisera) generation specific reaction, and be 0 with the HI reaction of H5, H7, H9 and ND positive serum;
(5) described strain has only EDS in duck embryo neutralization test
76Antiserum(antisera) can suppress the propagation of described strain virus at duck embryo allantoic cavity, and other positive serum all can not suppress the propagation of described strain virus;
(6) the relationship value R of described strain and international standard strain AV127 is 95%, belongs to same serotype;
(7) use the egg drop syndrome oil emulsion adjuvant emulsion of described strain preparation can stimulate chicken to produce EDS76 antibody.
3. the application of fowl egg drop syndrome virus stain as claimed in claim 1 in the production seed culture of viruses of preparation vaccine.
4. fowl egg drop syndrome virus stain as claimed in claim 1 is at preparation fowl egg drop syndrome viral diagnosis EDS
76Application in the antigen reagent.
5. the application of fowl egg drop syndrome virus stain as claimed in claim 1 in preparation fowl egg drop syndrome viral diagnosis positive serum reagent.
6. fowl egg drop syndrome virus stain as claimed in claim 1 is at preparation fowl egg drop syndrome viral therapy EDS
76Application in the antiserum(antisera) reagent.
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| CN 201010128150 Pending CN102086446A (en) | 2010-02-10 | 2010-02-10 | Strain of Adenoviridae and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102229916A (en) * | 2011-06-17 | 2011-11-02 | 福建省农业科学院畜牧兽医研究所 | Chicken Flavivirus and inactivated vaccine thereof |
| CN106177937A (en) * | 2016-07-25 | 2016-12-07 | 江苏省农业科学院 | Chicken egg drop syndrome and avian encephalomyelitis bivalent inactivated vaccine and preparation method thereof |
| CN117144065A (en) * | 2023-10-13 | 2023-12-01 | 河南农业大学 | Primer probe set, kit and detection method for fluorescence RAA detection of avian egg drop syndrome virus |
-
2010
- 2010-02-10 CN CN 201010128150 patent/CN102086446A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 《GENEBANK》 20071128 Avian adenovirus EDS gene encoding pVIII protein X99782.1 , * |
| 《中国畜禽传染病》 19940531 毛春生等 两株鸡减蛋综合征病毒在鸭胚中增值规律的观察 第78卷, 第5期 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102229916A (en) * | 2011-06-17 | 2011-11-02 | 福建省农业科学院畜牧兽医研究所 | Chicken Flavivirus and inactivated vaccine thereof |
| CN106177937A (en) * | 2016-07-25 | 2016-12-07 | 江苏省农业科学院 | Chicken egg drop syndrome and avian encephalomyelitis bivalent inactivated vaccine and preparation method thereof |
| CN117144065A (en) * | 2023-10-13 | 2023-12-01 | 河南农业大学 | Primer probe set, kit and detection method for fluorescence RAA detection of avian egg drop syndrome virus |
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