208 results about "Infectious bronchitis virus" patented technology

The invention discloses a GeXP (Gene Expression) rapid detection kit capable of simultaneously identifying six virus of chicken respiratory disease. The GeXP rapid detection kit is used basing on a CeXP system and contains seven PCR (Polymerase Chain Reaction) primer pairs, the specificity for simultaneously detecting avian influence virus, H5, H7 and H9 sub type of the avian influence virus, Newcastle disease virus, infectious bronchitis virus and infectious laryngotracheitis virus is strong, the sensitivity can be up to 100 copy / mu l, and compared with an identifying result of regular test methods such as virus isolation and hemagglutination inhibition, the coincidence rate is up to 100%. According to the GeXP rapid detection kit disclosed by the invention, a simple and high-throughput detection kit and a detection system are provided for the detection of common main chicken viral respiratory disease, the actual needs are accordant, and the application prospect is wide.

The invention provides an application of protocatechuic acid in the preparation of drugs for preventing and controlling livestock and poultry virus infectious diseases. The viruses include infectious bursal disease virus, avian influenza virus, infectious bronchitis virus and / or porcine transmissible gastroenteri tis virus. Meanwhile, the invention also relates to a protocatechuic acid preparation and a preparation method thereof.

The invention provides a preparation method for a triple inactivated vaccine of recombinant Newcastle disease, bird flu and infectious bronchitis. The technical scheme is realized by gene modification of a Newcastle disease virus (DNV) LaSota strain. A recombinant NDV low virulent strain (rLaSota strain) is taken as a vector, firstly an S1 gene of a chicken infectious bronchitis virus (IBV) is inserted between F-HN, a recombinant NDV rLaSota / S1 expressing an S1 protein is constructed by utilizing a reverse genetic technology, and a result shows that rLaSota can serve as a live vector for stably expressing the S1 protein; secondly a recombinant NDV rLaSota / S1-HA expressing an HA gene is constructed, namely, the HA gene is inserted between P-M of the recombinant NDV rLaSota / S1; and thirdly immunogenicity assessment is performed. A result shows that the triple inactivated vaccine of the recombinant Newcastle disease, the bird flu and the infectious bronchitis, prepared through a conventional inactivated vaccine process has relatively good protection effects for the Newcastle disease, the bird flu and the infectious bronchitis.

The invention relates to a method for producing a triple inactivated vaccine for newcastle disease, infectious bronchitis and infectious bursal disease. In the method, the inactivated vaccine is prepared by adopting a newcastle disease virus La Sota strain, an infectious bronchitis virus M41 strain, and an Escherichia coli genetic engineering bacteria E. coil BL21 / pET28a-VP2 strain which expresses an infectious bursal disease virus VP2 protein serving as production bacteria toxic species, a super concentration process and a high-quality adjuvant. After immunizing animals, antibodies are produced rapidly; the potency is high; the protection period is long; and the outbreak and the spreading of epidemic diseases are reduced.

The invention relates to a method for producing a combined vaccine, in particular to a method for producing a combined vaccine against Newcastle disease and infective bronchitis. The method comprises that: before culture in the same embryo, purity of breeding virus of the Newcastle disease and the infective bronchitis is improved; and during culturing in the same embryo, concentrations of two antigens in the combined vaccine against the Newcastle disease and the infective bronchitis are adjusted to reach reasonable combination. The vaccine prepared by the method is vaccinated with chicken flocks to generate ND and IB immunity without any interference. Furthermore, the vaccine strains are reasonably matched to effectively solve the problem of serious interference in simultaneous vaccination of the Newcastle disease living vaccine and infective bronchitis living vaccine, improve immunization quality and reduce the number of immunization.

The invention relates to a method for preparing a vaccine by breeding infectious bursal disease virus (IBDV) by a chicken embryo source cell line. The method mainly comprises the following steps of: 1) subculturing cells DF-1 for preparing vaccines; 2) breeding IBDV HQ cell seeds; 3) breeding virus liquid for preparing the vaccines; 4) concentrating and inactivating the virus liquid for preparingthe vaccine; 5) preparing other virus liquid of newcastle disease method, newcastle disease-infectious bronchitis virus method, and newcastle disease-infectious bronchitis virus-egg drop syndrome method combined vaccine and concentrating; and 6) proportioning inactivated combined vaccine, emulsifying and sub-packaging. The production process is simple, and stable and is easy to operate, eliminates biological potential safety hazard existing in the conventional vaccine production, and overcomes the defects that large-scale production of the vaccines is limited by supply of chicken embryos; cost and batch-to-batch variation are reduced; the virus titer and the quality of vaccine are improved; basis is laid for culturing virus liquid on large scale by a suspension culture technology in vaccine industry; and the produced IBDV inactivated vaccine and combined vaccine have high safety and immune efficacy, and have the complete immune protection effect on IBDV attack.

The invention is concerned with a kind of bacterin to prevent fowl infectious disease, relating to a kind of bacterin preparation method to prevent Newcastle Disease and infectious bronchitis. The selected genus is Newcastle Disease virus La Sota individual plant, infectious bronchitis virus M41 individual plant and HN99 kidney type of aberrance individual plant. Dilute two kind of virus and inoculate in chick embryo allantoic cavity, collect liquid of embryo to alive and dead embryos as seed. Dilute and inoculate in allantoic cavity of affectable chick embryo to get liquid of virus embryo. Concentrate with equipment and add with formaldehyde solution to prepare bacterin through emulsion process. This invention can prevent Newcastle Disease, chick kidney type and breathing type infectious bronchitis, and it can reduce the times of injection and the cost of epidemic prevention for easy using and practicality.

The invention provides a forage for preventing the infectious bronchitis of chicken. The forage is characterized in that the forage comprises corn, soybean meal, salt, fish meal, bone meal, a trace element additive, an amino acid additive, a vitamin additive, and a traditional Chinese medicinal immune powder feed additive used for preventing the infectious bronchitis of chicken. The forage has a substantial effect on the prevention of the infectious bronchitis of chicken, and has the advantages of non-toxicity, drug resistance and no drug residual.

The invention relates to a primer used in RT-PCR detection of chicken infectious bronchitis virus, a kit comprising the primer and a detection method thereof. The primer includes a specific primer pair used for amplifying a 5-terminal non-coding region conserved sequence of chicken infectious bronchitis virus (IBV). The specific primer pair is composed of a primer 1 and a primer 2, wherein the primer 1 is a single-chain DNA represented in the Seq ID No.1, the Seq ID No.2 or the Seq ID No.3 in the sequence table, and the primer 2 is a single-chain DNA represented in the Seq ID No.4, the Seq ID No.5 or the Seq ID No.6 in the sequence table. Tests comprising specificity, sensitivity, stability and repeatability prove that the kit can only specifically amplify IBV nucleic acid and is excellent in linearity relationship within the range of 1*10<2>-1*10<7> copies / [mu]L. The sensitivity of the primer reaches 10<2> copies / [mu]L. The primer is suitable for quantitative detection of various tissue, cell, body fluid samples and vaccine production intermediate products.

The invention relates to a method for producing a quadruple inactivated vaccine for newcastle disease, infectious bronchitis, avian influenza (H9 subtype) and infectious bursal disease. In the method, the inactivated vaccine is prepared by adopting a newcastle disease virus La Sota strain, an infectious bronchitis virus M41 strain, an avian influenza virus (H9 subtype) YBF003 strain, and an Escherichia coli genetic engineering bacteria E. coil BL21 / pET28a-VP2 strain which expresses an infectious bursal disease virus VP2 protein serving as production bacteria toxic species, a super concentration process and a high-quality adjuvant. After immunizing animals, antibodies are produced rapidly; the potency is high; the protection period is long; and the outbreak and the spreading of epidemic diseases are reduced.

The present invention belongs to the field of animal virology technology, and is especially one kind of short peptide capable of combining specifically with chicken infectious bronchitis virus (IBV) H52, bacteriophage containing the short peptide and its preparation process. The short peptide has the amino acid sequence of GSHHRHVHSPFV, and may combine with chicken infectious bronchitis virus H52 specifically after passing through the substitution, deletion or addition of one or several amino acid residues. The present invention also includes the application of the bacteriophage in preparing IBV identifying and diagnosing kit, and the IBV identifying and diagnosing kit is used in the preventing and treating of chicken IBV.

The invention discloses a visual protein chip for detecting serum antibody of new-castle disease virus of chickens, infectious bronchitis virus of chickens, avian influenza virus and infectious bursal disease virus of chickens , which is prepared by the following steps: purifying and diluting whole proteins of the four virus respectively; pointing samples of the positive control serum, the negative control serum and the four virus proteins onto a chip carrier respectively; drying, fixing, sealing and washing the samples to obtain the visual protein chip. The visual protein chip uses the purified whole proteins as capturing antigens to detect the virus-specific antibodies in chicken serum so as to simplify the preparation technology and reduce the production cost, and the visual protein chip has better specificity but no cross, has high reliability of results and has the advantages of quickness, simplicity and convenience, high sensitivity, good specificity and the like. When the serum is diluted by 6,400 times, the visual protein chip still can detect the antibodies, the sensitivity is 400 times of that of the prior AGP detection method. According to the detection to serum samples in-place, the detection rate of the visual protein chip is higher than the proir AGP method remarkably.

The invention discloses a recombination Newcastle vaccine strain rAI4-S1 for expressing infectious bronchitis virus S1 protein and a generating method of the recombination Newcastle vaccine strain rAI4-S1. The conservation number of the recombination Newcastle vaccine strain rAI4-S1 is CGMCC No: 8054. According to the generating method of the recombination Newcastle vaccine strain rAI4-S1, a built reverse genetic manipulation platform of a gene VII type NDV attenuated virus AI4 strain is utilized, the sequence revealed in the SEQ ID NO.7 is inserted into an AI4 strain genome full-length transcription carrier pNDV / AI4, and then the recombination Newcastle vaccine virus genome full-length cDNA clonal pNDV / AI4-S1 containing infectious bronchitis virus S1 genes is obtained. According to a recombination virus obtained through transfection, a chick embryo has high breeding titer, the S1 protein still can be expressed after continuous passage, and the generating method is suitable for large-scale production of vaccines and can be used for processing the vaccines.

Embodiments of the present invention generally relate to a novel attenuated infectious bronchitis virus (IBV) of GA-98 isolate. Further, other embodiments of the present invention generally relate to methods of immunizing avian against an infectious bronchitis virus. As well, further embodiments relate to method of making a vaccine and / or immunogenic composition for protecting avian, such as poultry, from an infectious bronchitis virus of strain GA-98.

The invention belongs to the field of the biotechnology and discloses an infectious bronchitis low-virulent live vaccine YX10 D90 strain. The infectious bronchitis low-virulent live vaccine YX10 D90 strain is preserved in China Center for Type Culture Collection on 14th March, 2016, the preservation number is CCTCC NO:V201610, and the preservation address is Wuhan University, Wuhan, China. The low-virulent strain is safe for all chickens and has no side reaction, the low-virulent strain is prepared into a vaccine safely and effectively, toxin attacking on homological strong toxicity can be protected, and practical and wide application value is achieved.

The invention is concerned with a kind of preparation method of inactivated vaccine to prevent new castle disease, infectious bronchitis and egg drop syndrome vaccine. The selected genus is Newcastle Disease virus La Sota individual plant, infectious bronchitis virus M41 individual plant, HN99 kidney type of aberrance individual plant and EDS76 Jing 911 individual plant. Dilute three kinds of virus and inoculate in the allantoic cavity of SPF chick embryo and affectable duck embryo. Collect liquid of embryo to alive and dead embryos as seed. Dilute and inoculate in allantoic cavity of affectable chick embryo or duck embryo to get liquid of virus embryo. Concentrate with equipment and add with formaldehyde solution to prepare bacterin through emulsion process. This invention can prevent Newcastle Disease, chick kidney type and breathing type infectious bronchitis, and egg drop syndrome vaccine. It can reduce the times of injection and the cost of epidemic prevention for easy using and practicality.

The invention belongs to the field of veterinary biological products, and relates to a strain of avian nephropathogenic infectious bronchitis virus containing a gene coding sequence SEQ ID No.2 which codes a protein and an attenuated virus strain. Serial passage of the virus strain is carried out to 110-140th generation in chicken embryos, and the attenuated virus strain is obtained. The invention also relates to a vaccine composition which is prepared by using the virus strain, the attenuated virus strain and antigen proteins thereof, as well as a preparation method and an application thereof. A vaccine composition containing inactivated totivirus antigen, live attenuated totivirus antigen and subunit antigen can be used for effectively preventing diseases related to avian nephropathogenic infectious bronchitis.

The invention discloses a method for quickly constructing an IBV (Avian Infectious Bronchitis Virus) reverse genetic strain, and belongs to the technical field of coronavirus reverse genetics. The constructing method comprises the following steps: quickly completing construction containing IBV genomic full-strength cDNA (Complementary Deoxyribose Nucleic Acid) clone by taking a BAC (Bacterial Artificial Chromosome) vector as a framework and applying an in-vitro homologous recombination technology, directly transfecting cells by a constructed recombinant plasmid, and transcribing in the cells, thus obtaining a transcript having infectivity; completing virus packaging; inoculating SPF (Specific Pathogen Free) chick embryo to a mixed solution of the cells and a culture medium and passing from generation to generation, thus obtaining the IBV reverse genetic strain. The constructing method disclosed by the invention has the advantages of simple operation and high positive cloning efficiency, the obtained IBV reverse genetic strain has passage stability, and an effective tool is provided for researching pathogenesis of the virus in vitro, developing a novel vaccine and the like; according to the method disclosed by the invention, transcription is carried out in the cells by utilizing a CMV (Cytomegalovirus) promoter added on a 5' terminal, and the rescue efficiency of the virus is greatly increased by utilizing an HDVR (Hepatitis Delta Virus Ribozyme) sequence added on a 3' terminal.

The invention provides a B cell antigen epitope polypeptide of a chicken infectious bronchitis virus Shanghai 1208 isolate S1 protein, and belongs to the field of gene and protein engineering. The B cell antigen epitope polypeptide can be combined to prepare an IBV multi-epitope vaccine. The IBV B cell antigen epitope can cause high-level immunoresponse.

The invention provides a method for production of an infectious bronchitis virus from a cell line. The employed cell line can be a Vero cell line, a BHK-21 cell line, a PK-15 cell line, a Marc145 cell line, a ST cell line, and an IBRS-2 cell line. The method includes: when the cell line forms a well-grown cell monolayer, inoculating an infectious bronchitis virus and making it adsorbed to the cell line; using a cell maintenance medium to conduct culture, adding an incubation agent to incubate the cell for 20-40min, and performing infectious bronchitis virus proliferation; and when CPE reaches over 75%, harvesting the infectious bronchitis virus. The method for production of an infectious bronchitis virus vaccine from a cell line provided in the invention can achieve the purpose of more secure and more effective infectious bronchitis virus and vaccine production.

The invention provides universal CD8+T cell epitope polypeptide of a chicken IBV (Infectious Bronchitis Virus) S1 protein, and belongs to the field of gene and protein engineering. The epitope polypeptides are prepared by the following steps: screening 21 epitope polypeptides in accordance with binding motif sequences in amino acid sequences of the S1 genes of the IBV virus according to the binding motif sequences of haplotype chicken major histocompatibility complex (MHC) I-type molecules; then, taking lymphocytes of three constructed SPF (Specific Pathogen Free) chicken immunized by DNA (Deoxyribonucleic Acid) recombinant plasmids of the S1 genes containing different subtype IBVs, determining the capacity of the 21 polypeptides which induce chicken splenic lymphocytes to secrete interferon-gamma by using an ELIspot (Enzyme-Linked Immunospot Assay) method, and finally, screening to obtain four universal functional T cell epitope polypeptides of IBV, wherein the sequences are respectively shown in SEQ ID NO. 1-4. The four epitope polypeptides provided by the invention are combined in use to prepare a universal vaccine for IBV. The invention further provides a method for screening the epitope of the functional T cell of the S1 protein of the IBV.

The invention discloses a poultry infectious bronchitis virus H120 reverse vaccine strain R-H120-Beaudette P65 (S) (Figure 1). The reverse vaccine strain is obtained by applying No See' M technology to replace S genes of an H120 strain with S genes of a PCR (polymerase chain reaction) amplification cell adapted strain Beaudette P65 strain. The reverse vaccine strain is adapted to Vero cells and can provide effective protecting force to chicken flocks. Through the reverse vaccine strain, the problem that IBV vaccine strains cannot be cultured in passage cells during production is solved, so that probability that vaccines are prone to being contaminated by exogenous viruses in the process of production is avoided, vaccine production quality is improved, and vaccine production cost is lowered greatly.

The invention discloses a method to sieve SARS coronavirus main prolease depressor from natural products mixture optional warehouse, which comprises the following steps: touching crystal of SARS coronavirus main prolease with 'sample with inhibited active'; getting electron density of small molecule depressant combined with SARS coronavirus main prolease; appraising the small molecule depressant based on electron density; getting fine three-dimensional structure of SARS coronavirus main prolease and small molecule depressant; getting small molecule depressant Sichuan scented sea vegetable ala factor. This Sichuan scented sea vegetable ala factor can inhibit coronavirus main prolease activator effectively, which can cure or prevent SARS coronavirus and infectivity gastrointestinal disease virus.

The invention relates to a production method of a newcastle disease (ND), infectious bronchitis (IB), infectious bursal disease virus (IBD) and viral arthritis (VA) four-joint inactivated vaccine. The production method is characterized by comprising the following steps: adopting ND La Sota strains, IB virus M41 strains, E.coli BL21 / pET28a-VP2 strains expressing the IBD virus VP2 protein and VA AV2311 strains as bacterial and viral strains and adopting a superstrong concentration process and high-quality adjuvants to prepare the inactivated vaccine. After being used for immunizing animals, the inactivated vaccine has the characteristics of high antibody production speed, high titer and long period of protection, and can be used for reducing the epidemic disease occurrence and spreading.

The invention provides a vaccine composition. The vaccine composition comprises an immune amount of an avian newcastle disease virus antigen, an avian infectious bronchitis virus antigen and an avian mycoplasma synoviae antigen and a carrier. The invention further provides a preparation method and application of the vaccine composition. The immune effect of the trigeminal vaccine is superior to the immune effect of combining newcastle disease-infectious bronchitis inactivated vaccines with mycoplasma synoviae inactivated single vaccine. The invention further provides a vaccine composition which contains a mycoplasma gallisepticum antigen in addition to the three antigens, and can be used for effectively resisting attack of four pathogens.

In the invention, a section of conserved sequences of four virus genes, namely Newcastle disease virus (NDV), infectious bronchitis virus (IBV), polymerase chain reaction (PCR) technology amplified egg drop syndrome (EDS) and infectious laryngotracheitis (ILT) by a molecular biotechnology (RT) PCR method is taken as a probe, a cellulose membrane is taken as a substrate (namely a most important basic platform in the detecting kit) to extract target pathogenic nucleic acid from pathological materials, and marked bio-16-dUTP is added during (RT) PCR to obtain a marked PCR product. The marked product is placed in the detecting kit for hybridizing detection, and quick and effective judgment and analysis on a biological sample are realized by observing changes of hybridization colors.

The invention discloses a dual reverse transcription-polymerase chain reaction (RT-PCR) detection method for identifying an H9N2 subtype avian influenza virus. A pair of primers is designed respectively according to HA genes and NA genes of the H9N2 subtype of avian influenza virus (AIV), the HA genes and NA genes of the H9N2 subtype AIV in a sample can be subjected to specific amplification, and lengths of target fragments are respectively 700bp (HA) and 423bp (NA). According to the method, cross reaction is avoided in subtype AIV such as H3N8, H4N6 and H5N8, Newcastle disease virus and infectious bronchitis virus of chicken; the lowest detection amount of allantoic fluid of the virus is 1*103.25EID50 / 100uL; compared with conventional methods such as hemagglutination inhibition of virus and a neuraminidase inhibition test, the method has the advantage that the coincidence rate of the identification result is 100 percent. A rapid, specific and sensitive detection means is provided for identifying the H9N2 subtype AIV. The detection method can be used for rapidly diagnosing diseases caused by the H9N2 subtype AIV and has good application prospects in aspects of clinical diagnosis and epidemiological investigation.

The invention provides a hybrid virus-like particle. The particle comprises a matrix protein M1 of influenza virus, surface antigen hemagglutinin (HA) of the influenza virus and fused membrane protein, wherein the fused membrane protein comprises an extracellular domain of major epitope of infectious bronchitis virus S protein, and a transmembrane domain and an intracellular domain of the HA of the influenza virus; the extracellular domain of the main surface antigen of the infectious bronchitis virus S protein replaces the extracellular domain with approximately same length of the 5'-terminal of the HA of the influenza virus; and the surface antigen HA of the influenza virus and the surface antigen S protein of the infectious bronchitis virus are expressed simultaneously on the surface of the hybrid virus-like particle. The invention also provides a bivalent vaccine of avian influenza and infectious bronchitis, and the bivalent vaccine comprises the hybrid virus-like particle and adjuvants. The invention also provides a preparation method of the hybrid virus-like particle.