160 results about "Marek's disease" patented technology

The present invention provides a novel avian herpesvirus (NAHV) vector and recombinant vaccines made therefrom that are useful to immunize avian species against Marek's disease, infectious laryngotracheitis and Newcastle disease. Methods of immunizing an avian species against Marek's disease, infectious laryngotracheitis and Newcastle disease are also provided.

The invention aims at providing an I-colony fowl adenovirus 4 strain, which is preserved with preservation number of CCTCC No. V201541. The I-colony fowl adenovirus 4 YBAV-4 strain disclosed by the invention is excellent in specificity and immunogenicity; a specific precipitation line does not appear in specific positive serum chicken SPF chicken serum such as infected cell sap and egg drop syndrome resisting virus, chicken infectious bursal disease virus, Newcastle disease virus, chicken infectious laryngotracheitis virus, chicken Marek's disease virus, avian influenza and the like, while an obvious specific precipitation line appears in I-colony fowl adenovirus 4 specific serum. The strain disclosed by the invention, as a vaccine strain which is good in manufacturing effect, is capable of preventing chicken hydropericardium syndrome, and the strain is applicable to identification of virus serotype and investigation on epidemiology.

Superior molecular vaccines comprise nucleic acids, including naked DNA and replicon RNA, that encode a fusion polypeptide that includes an antigenic peptide or polypeptide against which an immune response is desired. Fused to the antigenic peptide is an intercellular spreading protein, in particular a herpes virus protein VP22 or a homologue or functional derivative thereof. Preferred spreading proteins are VP22 from HSV-1 and Marek's disease virus. The nucleic acid can encode any antigenic epitope of interest, preferably an epitope that is processed and presented by MHC class I proteins. Antigens of pathogenic organisms and cells such as tumor cells are preferred. Vaccines comprising HPV-16 E7 oncoprotein are exemplified. Also disclosed are methods of using the vaccines to induce heightened T cell mediated immunity, in particular by cytotoxic T lymphocytes, leading to protection from or treatment of a tumor.

The present invention discloses novel recombinant multivalent non-pathogenic Marek's Disease virus constructs that encode and express both Infectious Laryngotracheitis Virus and Newcastle Disease virus protein antigens, and methods of their use in poultry vaccines.

The invention provides a GeXP quick detection kit and detection method for simultaneously identifying 8 chicken infectious immunosuppression disease pathogens. The kit is used on the basis of a GeXP system, and comprises 8 pairs of PCR (polymerase chain reaction) primers. The detection result indicates that the kit has the advantages of high specificity and high sensitivity and can be used for simultaneously identifying and detecting chicken Marek's disease virus, A, B and J subgroup avian leucovirus, avian reticuloendothelium hyperplasia virus, avian reovirus, chicken infectious anemia virus and infectious bursal disease virus.

The invention relates to a construction and an application of recombinant Chicken Marek's Disease Virus SC9-1 strain and SC9-2 strain. The recombinant virus construction method solves the technical problem that kanr gene cannot be knocked out again by a present method after kanr gene containing flp recognition sites at two ends is continuously used twice to knock out two specific functional genes on the same virus genome. The obtained recombinant virus MDV SC 9-1 strain and MDV SC 9-2 strain are used as production strains of Marek's Disease live vaccine. The prepared vaccine prevents the very virulent or emerging very virulent plus MDV induced chicken Marek's disease. The protective immunity effect of the vaccine is superior to CVI988 / Rispens strain vaccine which is mostly widely used in foreign and domestic markets at present. The antigenicity of the recombinant virus is more similar to that of Chinese epidemic strain than the antigenicity of the similar virus rMd5deltameq which has been published in the United State. The strains provided by the invention will not induce tumor and has no immune suppression effect. Therefore, the strains are more applicable to China.

The invention discloses a chicken Marek's disease (MD) Meq gene deleted vaccine strain, a construction method thereof, and an application thereof. The chicken Marek's disease rMS delta Meq gene deleted vaccine strain has a microbe reservation number of CGMCC No. 4612. According to the invention, on a basis of a parent strain which is MDV MS strain, a 468bp base sequence on a front part of a main oncogene is deleted thorough two times of homologous recombination, such that the chicken Marek's disease Meq gene deleted vaccine strain is obtained. The invention also relates to the application of the gene deleted vaccine strain in preparation of medicine used for controlling chicken Marek's disease, and in detection and disgnosis methods used for distinguishing a vaccine strain and a chicken Marek's disease wild strain, wherein the methods are designed aiming at the deleted gene sequence of the gene deleted strain. The gene deleted vaccine strain provided by the invention has good safety and good immuno-protection effect upon chicken Marek's diseases. The vaccine strain can be used for preparing monovalent vaccines or combined vaccines used for controlling chicken Marek's disease.

The invention discloses a chicken Marek's disease virus (MDV) miRNA (micro Ribonucleic Acid) deletion vaccine strain and application thereof. The invention is characterized in that: a recombinant strain rMSdeltamiR9-12 strain is constructed by taking a recombinant MDV circulating strain rMS-LacZdeltaMeq which carries a beta-galactosidase reporter gene and deletes a Meq gene as a parent strain, and deleting mdv-miR-M9, mdv-miR-M5 and mdv-miR-M12. The invention further discloses the application of the gene deletion vaccine strain to the prevention and treatment of the chicken MDVs and the application effect. The microbial collection number of the gene deletion vaccine strain is CGMCC No.4613. The miRNA deletion vaccine strain has stable heredity and high safety, has a good immune protection effect on the chicken Marek's disease, and can be prepared into a single vaccine or a combined vaccine and the like for preventing and treating the chicken MDVs.

The invention provides a method for labeling antibodies by colorful fluorescent granules and a test paper strip prepared from the antibodies. Animal viruses including, but not limited to, canine distemper viruses, canine parvoviruses, canine adenoviruses, canine coronavirus, rabies viruses, feline leukemia viruses, Marek's disease viruses, Newcastle disease viruses and the like are used as targets, and the corresponding antibodies labeled by colorful fluorescent micro-spheres are prepared by the aid of chemical covalent processes and can be applied to detecting the animal viruses. The method for labeling the antibodies by the colorful fluorescent granules and the test paper strip prepared from the antibodies have the advantages that colorful detection lines with bright developed colors can appear during detection, and accordingly qualitative results can be obtained; the contents of the animal viruses in samples further can be subsequently quantitatively obtained, accordingly, test results are clear and are high in stability, and the test paper strip can be stored at the room temperature for a long time.

The invention discloses a multiple fluoroimmunoassay primer, kit and method for rapidly distinguishing 5 avian immunosuppression pathogens. Operation is simple, a target amplified fragment is acquired through a PCR, an amplified product, fluorescence coded microspheres and streptavidin-phycoerythrin are hybridized, the MFI value is read through a detector, and viruses of different types are distinguished. The method can simultaneously detect the chicken infectious anemia virus, the chicken Marek's disease virus, the avian reticuloendotheliosis virus, the avian reovirus and the chicken infectious bursa disease virus accurately, specificity is high, sensitivity is high, and repeatability is good. Compared with a traditional detection method, the method achieves simultaneous detection of multiple different target molecules in the same sample, the use number of samples is small, operation is simple and rapid, and the detection cost can be greatly reduced.

The invention discloses a recombined chicken Marek's disease virus vaccine strain for expressing an infectious bursaldisease virus VP2 gene and a construction method and application of the recombined chicken Marek's disease virus vaccine strain, and belongs to the technical field of medicine or veterinary medicine. By means of the recombination and clone technology, a gene segment CAG-VP2 containing the infectious bursaldisease virus VP2 gene and a CAG promoter sequence is inserted in a US2 gene of the strain 814 of the chicken Marek's disease virus, a recombined cosmid with a CAG-VP2 expression frame inserted in a US2 gene is constructed, and the recombined chicken Marek's disease virus vaccine strain for expressing the infectious bursaldisease virus VP2 gene is obtained through salvation of the recombined cosmid. Research shows that the obtained vaccine strain has the same in-vitro replication ability as a parent virulent vaccine strain 814 and good hereditary stability, and can resist attacks of a very virulent MDV strain and a very virulent IBDV strain at the same time. It can be seen that the obtained recombined MDV vaccine strain can be used for preparing medicine for preventing or treating infectious bursaldisease and the chicken Marek's disease.

The invention belongs to the technical field of biological medicament making, and discloses a method for preparing a Marek's disease liquid nitrogen vaccine (814-strain) by utilizing a cell factory and the Marek's disease liquid nitrogen vaccine (814-strain) prepared by the method. The Marek's disease liquid nitrogen vaccine (814-strain) produced by utilizing the cell factory has high quality, a high immune efficiency and a stable immune effect; in addition, the Marek's disease liquid nitrogen vaccine also has the advantages of low generation, small plaque, high plaque forming unit (PFU) content, high immunogenicity, low cost, capacity of generating immunoprotection in a relatively short time and the like.

The invention relates to bird flu and Marek's disease dual live vaccine rMDV-HA viral strain and a construction method. H5 subtype bird flu virus SY strain haemagglutinin genes, reticuloendotheliosis disease virus LTR, screening mark genes lac / smGFP and Marek's virus Rispens CVI 988 vaccine strain genome exogenous genes are amplified by adopting reverse transcriptase-polymerase chain reaction or polymerase chain reaction and cloned to plasmid vectors to form recombinant plasmids pHA, pLTR, ploxP-lac / smGFP-loxP, pUP and pDOWN; DNA of transfer vector pMHA plasmids and DNA extracted from cells infected from the Marek's virus Rispens CVI 988 vaccine strain are co-transfected to chick embryo fibroblast; the exogenous genes are inserted to the Marek's virus Rispens CVI 988 vaccine strain genome through homologous recombination to form recombinant virus rMDV-HA / GFP with the screening mark genes; and the screening mark genes lac / smGFP of the rMDV-HA / GFP are removed through cre enzyme-mediated loxP site sequence recombination, and the rMDV-HA / GFP is purified to form the rMDV-HA viral strain. The rMDV-HA viral strain has the advantages of low cost, no pollution and long immunity period, and can be used as bird flu and Marek's disease dual live vaccine viral strain.

The invention discloses a method for detecting Marek's disease resistance homozygous genotype chicken, which comprises the following steps: adopting the PCR-RFLP technology to perform PCR amplification on the second exon of BLB2 genes of 99 MDV-1 toxin eliminated Xiayan chickens and perform Alu I, Cai I, Cfr I, Hinl I, Hinf I and Rsa I restriction enzyme analysis on the PCR products, and initially obtaining seven BLB2 allelic genes genotype chickens by performing the restriction analysis on the six loci; finally obtaining six BLB2 / BF2 allelic genes genotype chickens by sequencing the BLB2 and BF2 genes of the seven genotype chickens; and by combining the toxin-eliminating result, determining four MD resistance individuals. The established method can detect the MD resistance homozygous genotype chickens, and the MD resistance homozygous genotype chickens are taken as breeders to breed the MD resistance chickens. Therefore, the method has extremely great economic benefits to prevention and control of MD and related immune-suppressing diseases on genetic predisposition.

The invention relates fodder special for breeding for improving disease resistance of peacocks. 1 kg of the fodder comprises 200-300 g of oat, 80-100 g of sorghum, 20-40 g of seaweed seeds, 3800-3900 mg of lactic acid, 100-112 mg of glucose oxidase, 132-138 mg of calcium formate, 6-5 g of mirabilite, 10-12 g of magnolia flowers, 8-10 g of royal paulowinia flowers, 3-5 g of mentha crispate, 2-3 g of oriental wormwood, 5-8 g of radices trichosanthis, 6-10 g of felwort, 2-3 g of white mustard seeds, 2-5 g of aristolochia debilis, 10-15 g of platycodon grandiflorus and the balance corns. Traditional Chinese medicine fodder components such as the oriental wormwood, the radices trichosanthis, the felwort, the white mustard seeds, the aristolochia debilis and the platycodon grandiflorus are added to the fodder so that the fodder can have the obvious function of improving immunity of the peacocks, the traditional Chinese medicine fodder components can be matched with the rest of the feed raw materials to achieve the effects of regulating qi and blood, driving out evil spirits and clearing heat, resisting bacterium and diminishing inflammation, expelling insects and removing qi stagnation and building and activating the spleen and the stomach, a good preventing effect is achieved on enteritis, marek's disease and coccidiosis occurring in peacock breeding, and a good effect is achieved on improving the production performance of the peacocks, protecting the health and promoting growth.

The present invention provides a novel avian herpesvirus (NAHV) vector and recombinant vaccines made therefrom that are useful to immunize avian species against Marek's disease, infectious laryngotracheitis and Newcastle disease. Methods of immunizing an avian species against Marek's disease, infectious laryngotracheitis and Newcastle disease are also provided.

The invention discloses a making method of live polyvalent vaccine in the molecular biological domain, which protects five virus infections within one time through sucking and / or intaking expressive infectious bronchitis, marek's disease, facystic disease, newcastle disease and poultry influenza virus.

The invention discloses a production method for a chicken Marek's disease vaccine by using a continuous passage cell line. The method comprises the following steps: (1) passage and culture of a cell used for vaccine preparation; (2) propagation of a cellular virus seed; (3) inoculation of a virus; (4) centrifugal extraction of the cell for collection of the virus; and (5) distribution and split charging of the vaccine. According to the invention, passage cells are used to replace original primary chicken embryonic fibroblasts for production of the chicken Marek's disease vaccine, so production cost is substantially reduced; the production method has a simple process, is stable and is easy to operate; virus plaque content is high, the vaccines of different batches have small difference, vaccine quality is easily controllable, and output and quality of the vaccine can be substantially improved. The chicken Marek's disease vaccine produced in the invention has high immune efficacy on a standard virulent strain and has an immune effect superior to that of a product of a same kind.

The present invention provides an active vaccine rHVT-HA for prevention and treatment of avian influenza and Marek's disease. The HVT is recombined with a HA gene of avian influenza virus, and the HA gene can express HA protein. The live vaccine can rapidly generate HI antibody in the body, has more effective prevention effect on avian influenza effect than the traditional inactivated vaccine, and has good protective effect on the infection of Marek's virus.

The invention discloses a multiplex detection reagent for immunosuppressive poultry pathogens of avian influenza and the like and an application of the multiplex detection reagent. The multiplex detection reagent comprises six specific probes with sequences shown in Seq ID NO.1-Seq ID NO.6 which refer to AVI virus, ND (newcastle disease) virus, serotype I Marek's disease virus, IBDV (infectious bursal disease virus), avian infectious anaemia virus and ALV-J (aivan leukosis virus subgroup J) in sequence. The reagent comprises the specific probes for six immunosuppressive poultry pathogens comprising AVI and the like and can specifically capture target sequences of the six immunosuppressive poultry pathogens, and the foundation is laid for high-throughput rapid detection of the six immunosuppressive poultry pathogens. With adoption of the reagent, a rapid and accurate detection method is provided for the six immunosuppressive poultry pathogens on the basis of a liquid chip.

The present invention provides an avian recombinant herpesvirus modified by the presence of the cDNA encoding, the VP2 of the Delaware Variant E strain of IBDV, a subtype of IBDV serotype 1 strains. The present invention further provides an avian recombinant herpesvirus comprised of the VP2 gene of which the backbone virus is a Marek's disease vaccine strain, such as herpesvirus of turkeys. A poultry vaccine including the avian herpes recombinant virus described in the present invention can induce in chickens protective immunity against a variety of different subtypes of IBDV.

The present invention provides an effective vaccine for Marek's disease, which may be prepared using a recombinant Marek's Disease Virus (MDV), strain CVI988, having been transformed with a foreign DNA construct that includes a long terminal repeat sequence of a reticuloendotheliosis virus. This safe viral agent elicits a highly protective immune response in a chicken against virulent MDV challenge without causing a significant degree of pathogenicity. Suitable formulations of the vaccine for use in chickens include an effective immunization dosage of this novel viral agent, along with a pharmaceutically acceptable carrier or diluent.

The invention belongs to the technical field of biopharming, and discloses a method for preparing a chicken Marek's disease turkey herpes virus Fc126 strain live vaccine by utilizing a cell factory and the turkey herpes virus vaccine (Fc126 strain) prepared by utilizing the method. The turkey herpes virus vaccine (Fc126 strain) prepared in the cell factory is good in quality, high in immunization potency and stable in immunization effect. In addition, the turkey herpes virus vaccine (Fc126 strain) has the advantages of being low in generation, high in content of plaque forming unit (PFU), low in cost, simple in operation technology, easy to control and the like.

The invention discloses a preparation method of AGP detection antigen used for diagnosing Marek's virus infection, relating to the poultry disease detection reagent field. The method comprises the steps as follows: firstly, the separating effect experimental evaluation is carried out on the target virus antigen, so that a permeation-typed filter membrane element and a retention-typed filter membrane element are screened out from ultrafiltration membrane elements with different molecular weight cutoff; secondly, the permeation-typed filter membrane element and the retention-typed filter membrane element which are screened out are respectively used to purify and concentrate the liquid containing the target virus antigen; finally, the purified and concentrated solution is detected and quantified, so that the Marek's virus infection AGP detection antigen is prepared; and no reagent is added in the process of ultrafiltration treatment, therefore, not only neither secondary pollution nor contamination is produced in the production process, but also the emission liquid is actually further purified, so as to reduce the pollution to the environment and improve resource utilization, thereby really achieving the purpose of transforming trash into treasure, and the preparation method also has the advantages of no phase state change, mild conditions, simple operation, low cost and high recovery rate, etc.

The invention discloses a method for culturing directly differentiated tissues of a cotton true leaf and a special culture medium. The special culture medium provided by the invention is a culture medium which is used when embryogenic callus is directly differentiated from a cotton leaf; and the culture medium is obtained by adding KT, 6-BA, 2,4-D, indole acetic acid (IAA), glucose and gel into marek's disease 1 (MSB1) basic culture solution, wherein the final concentration of the KT, the 6-BA, the 2,4-D, the IAA, the glucose and the Gel rite is 0.005 to 0.15mg / L, 0.005 to 0.15mg / L, 0.0001 to 0.001mg / L, 0.001 to 0.02mg / L, 25 to 35g / L and 1.8 to 2.5g / L respectively. By the method, a tissue culture system is established by taking leaves of field cotton plants as explants; and finally, a stable and high-efficiency tissue culture system of the directly induced embryogenic callus of the leaf is established.

The invention belongs to the technical field of biology and in particular discloses a recombinant herpesvirus of turkey (HVT). The recombinant HVT is capable of simultaneously expressing NDV and IBDVprotective antigen genes, and is a bacterial artificial chromosome (BAC) based on the HVT. The construction method comprises the following steps: inserting a VP2 gene expression cassette of an infectious bursal disease virus LX strain under regulation of a cytomegalovirus promoter CMV into a UL45-46 non-essential region of the HVT genome by utilizing a gene recombination technology; and insertingan F gene expression cassette of a newcastle diseases virus PX02 / 3 strain under regulation of a chicken beta-actin promoter pec into another non-essential region US1-10 of the HVT genome. The recombinant herpesvirus of turkey is a recombinant triplet poultry vaccine candidate strain capable of controlling Marek's disease of chickens and effectively resisting Newcastle disease and infectious bursaldisease.

Provided is a method for producing a continuous cell line capable of supporting the growth of a Marek's disease virus (MDV) strain comprising infecting or transfecting a cell with a vector comprising a nucleic acid or fragment thereof of an MDV strain and culturing the cell or a progeny thereof under conditions suitable for the expression of the nucleic acid and propagation of the cell or progeny. The nucleic acid may comprise an MDV glycoprotein gE gene or functional fragment. Also provided is a method of generating, isolating, and / or maintaining a hypervirulent, very virulent, very virulent plus, virulent, and / or avirulent MDV strain comprising infecting a described cell with the MDV strain or strains, and culturing the cell under conditions suitable for the propagation of the cell and the generation, maintenance and isolation of the MDV strain or strains. Also provided is a method to prepare a vaccine capable of inducing protection against disease, preferably an MDV associated disease in an avian, including culturing a continuous cell according to the invention and harvesting cell culture components therefrom.

The invention discloses a reverse genetic operating system for a Marek's disease virus attenuated vaccine strain 814 and an application thereof in virus rescue. A genome of the Marek's disease virus attenuated vaccine strain 814 is cut and then subjected to segmented cloning into a Fosmid cosmid vector pCC1Fos, and a Fosmid library of the genome of the vaccine strain 814 is constructed. On the basis of genome sequencing on the library, 5 recombinant cosmids which are cloned with the vaccine strain 814 genome segment, mutually contain overlapping regions and can be spliced to cover the whole genome of the strain 814 are selected. The 5 cosmids are used for co-transfecting chicken embryo fibroblasts (CEF), and viruses having same biological characteristics with an original MDV vaccine strain are successfully rescued. With the application of the established reverse genetic operating system, firstly, the attenuated vaccine strain can be rescued and used as a vaccine for MDV prevention; and more importantly, with the use of the reverse genetic operating system, virus transformation can be implemented in bacteria, so as to be used in development of MDV live vector vaccines.

The invention discloses a multiple fluorescence immunoassay primer, kit and method capable of fast distinguishing CAV, MDV, REV and IBDV. The multiple fluorescence immunoassay primer, kit and method has the advantages that target amplification fragments are obtained through PCR, \ amplification products, fluorescence encoded microspheres and streptavidin-phycoerythrin are hybridized, MFI values are read through a detector, and different types of pathogens are distinguished; the method can simultaneously and accurately detect chicken infectious anemia viruses, chicken Marek's disease viruses, chicken reticuloendotheliosis viruses and chicken infectious bursal disease viruses and is high in specificity, high in sensitivity and good in repeatability; compared with a traditional method, the method can simultaneously detect various molecules, with different purposes, in the same sample and is low in sample use amount, simple and fast to operate, capable of greatly lowering detecting cost, good in flexibility and capable of increasing and decreasing to-be-detected pathogen types on the basis.