185 results about "Animal virus" patented technology

The invention relates to the technical field of cell cryopreservation liquids, and in particular relates to a cryopreservation liquid for cultured NK cells and a preparation method of the cryopreservation liquid. The cryopreservation liquid for the cultured NK cells comprises tea polyphenol, a grape seed extract, vitamin C and autoserum. By virtue of adopting the autoserum, pollution of exogenous animal viruses and introduction of exogenous proteins are avoided, and the safety is high; by virtue of adding the tea polyphenol, the grape seed extract (procyanidine) and the vitamin C, cell oxidization can be effectively suppressed, the activity of the cells can be retained, and no harm to a human body is generated; meanwhile, the state of the cells recovered from cryopreservation is basically close to the state before cryopreservation, and the cryopreservation effect is good.

The present invention relates to the field of animal virology, and provides a recombinant chicken-origin H9N2 avian influenza virus vaccine strain and a method for isolation, identification and purification of the strain. The invention further relates to a research of biological characteristics of the strain, especially to a research of characteristics of the strain adopted as the vaccine strain,and an evaluation of immune effects of the strain on SPF chickens. The preservation number of the strain is CCTCCNO:V201030. According to the present invention, the antigen variation conditions of the virus strain and other isolated virus strains are represented from the molecular level; after the virus strain is prepared into the vaccine, the prepared vaccine is adopted to immunize the 4 week old SPF chickens, with the protection effect analysis of the homologous H9 influenza wild virus strain and the heterologous H9 influenza wild virus strain, the results show that the influenza virus strain can be adopted as the spare vaccine strain of H9 subtype avian influenza. With the present invention, the spare vaccine strain is provided for prevention of the avian influenza outbreak by using the vaccine, the molecular biology technology program is provided for screen of the avian influenza virus vaccine strain, the molecular biology background is provided for study of the mechanism of animal infection by the avian influenza, and the important public health significance is provided.

The invention discloses an on-site detection immuno-chip, comprising a chip carrier and an agarose gel layer which is paved on the chip carrier, wherein, the agarose gel layer is fixed with a plurality of antibody microarrays of 4*4, and chip-dedicated fences or Super PAP Pen scribings are used for separating the microarrays from each other; the double antibody sandwich principle is adopted to detect the antigen to carry out the dot matrix of corresponding capture antibody, positive control and negative control on the solid phase carrier simultaneously; the antibody protein is connected with the solid phase carrier through the covalent bond and physical adsorption; the sample liquid to be detected and the chip are directly incubated; the antigen to be detected in the samples combine with the corresponding antibody which is fixed on the chip; a specific monoclonal antibody probe marked by horse radish peroxidase is added and the macroscopic detection results can be obtained after the coloration of substrates. The invention can detect viruses in multiple aquatic animal samples and the results are macroscopic, thus being applicable to rapid and accurate detection of viruses of aquatic animals in breeding production.

The invention discloses two types of ScFv (Single Chain Variable Fragment) antibodies, encoding genes of the antibodies and applications of the antibodies for preparing the preparations for treating or preventing infectious bursal disease of chicken. Two single-chain antibodies (namely, the ScFv antibodies) are provided; and the ScFv antibodies can be specially bound with the protein 2 (VP2) (Virus Protein 2) in an IBDV (Infectious Bursal Disease Virus) structure and a plurality of IBDV strains to block the cytopathic effect (CPE) of the chicken embryo fibroblast by the IBDV so as to protect the young chicken infected with IBDV. The immune serum and egg yolk antibody are tedious in preparation, high in production cost, unstable in effect, difficult to control industrialized production quality, capable of causing horizontal disease spread and the like in a use process; the ScFv antibodies disclosed by the invention can overcome the above disadvantages and have the advantages of strong specificity, good treatment effect, controllable industrialized production quality, capability of preventing the horizontal disease spread caused by the egg yolk antibody and the like; and therefore, by virtue of the ScFv antibodies, a new situation is opened up in the IBDV prevention and treatment history, and even in the entire prevention and treatment history for animal virus diseases.

The invention provides a human McAb to tetanus toxin, which is a human antibody comprising a variable region of the antibody. The antibody consists of or not consists of a constant region of the antibody and has the ability of neutralizing the tetanus toxin, with the variable region gene and protein sequence indicated as the sequence table 1. The antibody is characterized in that the antibody can not induce obvious allergic reaction, has higher titer and longer effect without any animal virus pollution, and can be produced in an unlimited quantity. The antibody also provides the relating biological functions, the testing methods, the manufacturing method and the application.

The invention discloses a colloidal gold chemiluminescence immune assay method of detecting pig peripneumonia antibody. The method relates to animal virus immunodetection and chemiluminescence immune assay field. The main points of the invention are hereinafter: fixing the pig peripneumonia Apx IVA antigen in 96-microtitor plate, blocking the spare active site in the plate, add sample product to be examined, thus the pig peripneumonia Apx IVA antigen is binded. The gold-labeled rabbit anti pig IgG is added and the specific reaction with the sample can take place. The chemiluminescence reagent is added with the dissolved solution after the colloidal gold labeled on rabbit anti pig IgG is dissolved by violet acid and then detect chemiluminescence intensity. The chemiluminescence intensity is in direct ratiot to antibody concentration to be detected, thus linear correlation is available between chemiluminescence intensity and pig peripneumonia Apx IVA antibody. Compared with the prior art, the method has more accuracy and high specificity to better meet the need of clinical detection of peripneumonia.

The invention provides a method for enriching viruses from faeces rapidly and efficiently, amplifying signals through gold nanoparticles crosslinked through specific DNA labels and then rapidly detecting early-state PEDV infection with a PCR. According to the method, an PRF1a gene located in a PEDV whole genome is adopted as a target sequence, specific nucleic acid probes are designed, second-segment probes which are high in specificity, great in virus enriching capability and easy to amplify and detect are screened out of the specific nucleic acid probes and optimized, and by means of functionalized magnetic particles and nanogold particles of the second-segment probes, a PCR detection technology based on nanoparticle PEDV specificity is established; meanwhile, by means of the technical method, the technical breakthrough of conducting a direct hybridization reaction between faeces sample lysate and the functionalized magnetic particles is achieved, the step of purifying a sample nucleic acid is completely omitted, specificity and sensitiveness are improved, meanwhile, the detection step is simplified, and detection time is shortened. By means of the method, time and labor are saved, cost is low, and in addition, the animal virus infection level can be prompted accurately.

The invention relates to the technical field of animal virus detection in the field of veterinarians, and particularly discloses a real-time fluorescent PCR primer probe combination and a kit for detecting African swine fever virus wild strains. A PCR method based on the kit can not only specifically detect the African swine fever virus wild strains, but also serve as a method for distinguishing between the African swine fever virus wild strains and MGF360-505R loss attenuated vaccine virus strains when in use in combination with current p72 fluorescent PCR, therefore, important tools are provided for detecting the African swine fever virus wild strains and animals infected by the African swine fever virus wild strains, and control over African swine fevers and purification of the Africanswine fevers are promoted.

Based on a highly conserved domain of a foot-and-mouth disease virus 3D protein coding gene, a vesicular stomatitis virus N protein coding gene and a swine vesicular disease virus VP1 protein coding gene, the invention designs a specific primer and a probe and develops a multiple fluorescence RT-PCR detection method used for simultaneously detecting the three animal viruses. The detection method can detect 102 copied plasmids containing target amplification sequences in 1 h. The method has high sensitivity and good specificity, and can achieve rapid high-throughput fluorescence RT-PCR detection for single-virus infection or mixed infection by a plurality of viruses.

The invention belongs to the technical field of animal virology and animal-borne disease, and particularly relates to a low virulent strain variated from the porcine epidemic diarrhea virus and application thereof. The low virulent strain is a porcine epidemic diarrhea virus AJ1102-R strain having the preservation number of CCTCC NO:V201433, and has stable breeding performance after verification of cell passage and plaque cloning and culturing. The genomic sequence of the low virulent strain has the following variations: the 96th generation has sequence variation on the 71-72 of 3'UTR, and 2 bases TT are lost; and the 96th generation has sequence variation on the 3805-3816 of the S gene, and 12 bases GATGCTATTGTT are added. The immune efficiency of the low virulent strain can be used for inducing an immunized piglet to generate high-level neutralizing antibody and enough attacking protection, and can be used as a vaccine strain for application.

The invention relates to a grass goldfish pelvic fin cell line and a constructing method thereof. Tissue blocks are digested through trypsin, EDTA and hyaluronidase according to the features of grass goldfish pelvic fin tissue histocyte, the digested tissue blocks are transplanted to a cultivation bottle, special multiplication cultivation solution is added to the cultivation bottle, primary culture is conducted, and then the grass goldfish pelvic fin cell line is successfully constructed. By means of the method, a large number of cells can be separated three days after primary culture is conducted, and the cells have the continuous passage capability. After passage is stable, the cells are in a typical fiber form, it proves that the Chinese grass goldfish pelvic fin cell line is stable in character and uniform in component and is a good material for studying the aquatic animal virology and the immunology. At present, the grass goldfish pelvic fin cell line has 60 generations, and the cells can stably increase and have the continuous passage capability.

The application provides a feed additive, which comprises higher-level fatty acid monoglyceride. The application also provides application of the feed additive in the preparation of a feed for controlling animal enveloped virus. The application also provides a feed containing the above feed additive. The feed additive contains higher-level fatty acid monoglyceride with low toxicity, has a good effect of controlling enveloped virus, and is a feed additive for wide non-specific control of animal virus. Investment in manpower is not required, stress reaction on animals will not be caused and growth of animals will not be affected, and growth of animals will be promoted. It shows through test results that loss of catastrophic virus infection of farmed animals can be reduced by at least 50% by the use of the feed additive.

The invention discloses a scFv antibody, an encoding gene thereof and application thereof to preparation of preparations for treating or preventing an infectious bursal disease. The scFv antibody is formed by a heavy chain variable region, a light chain variable region and a connecting region between the heavy chain variable region and the light chain variable region. The heavy chain variable region is (a) or (b), wherein (a) refers to protein which is formed by amino acid residues at the first site to the 124th site of the sequence 1, and (b) refers to protein which is subject to substitution and/or depletion and/or addition of the amino acid residues of (a) and has the same activity; and the light chain variable region is (c) or (d), wherein (c) refers to protein which is formed by amino acid residues at the 140th site to the 244th site of the sequence 1, and (d) refers to protein which is subject to substitution and/or depletion and/or addition of the amino acid residues of (c) and has the same activity. The scFv antibody provided by the invention has the advantages of high specificity, good curative effect and the like and creates a new situation in the prevention and treatment history of IBDV (Infectious Bursal Disease Virus), and even in the prevention and treatment history of the entire animal virus diseases.

The present invention is directed to compositions and methods for the induction of immune responses in mammals against enveloped animal viruses. More particularly, the invention provides vaccine compositions containing multiple MHC allotypes. By generating an immune response against these MHC molecules, virus or virus-infected cells expressing foreign MHC molecules can be attacked prior to infection of cells in the immunized host. In some embodiments, the vaccine compositions contain viral antigens and adjuvants as well. The vaccine compositions may comprise intact cells, cell-derived membrane preparations or recombinantly or chemically produced MHC molecules or fragments thereof.

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 relates to the technical field of cell cryopreservation solutions, and in particular relates to a cryopreservation solution for the cultured NKT cells and a preparation method of the cryopreservation solution. The cryopreservation solution for the cultured NKT cells comprises lentinan, algal polysaccharides, autoserum and DMSO. By virtue of adding the autoserum, the pollution of foreign animal viruses and the introduction of foreign proteins are avoided, and the safety is high; by virtue of adding the lentinan and the algal polysaccharides, the activity of the cells can be effectively retained, and no harm to a human body is generated; meanwhile, the state of the cells recovered from cryopreservation is basically close to the state before cryopreservation, and the cryopreservation effect is good.

A monoclonal antibody-based Dot-ELISA assay for the rapid detection of animal viruses such as avian influenza virus. The assay includes applying a specimen suspected of containing an animal virus on a porous membrane and treating the specimen with a solution of citric acid or lactic acid and a solution containing a mucolytic agent and a detergent. The treated specimen is then contacted with a primary monoclonal antibody for detecting the virus. If present, the primary moncolonal antibody bind with an antigen of the animal virus specimen. The specimen is contacted with an anti-monoclonal antibody conjugate (secondary antibody) and incubated to facilitate binding of the antigen-bound monoclonal antibody to the conjugate. The bound conjugate and antigen-bound monoclonal antibody is contacted with a coloring reagent to allow visual detection of the presence of the animal virus in the specimen.

The invention discloses the cDNA sequences of the non-structural protein gene NS1 of avian influenza virus, its preparation process and use, wherein gene is employed for constructing prokaryotic expression vectors, the expression vectors are transformed into bacillus coli to obtain recombinant strains (Escherichia coli BL21/pET-28a-NS1), the strains are utilized to express the non-structural protein NS1 of the avian influenza virus, the expression proteins are used to establish a differential diagnosis method through enzyme linked immunosorbent assay for differentiate vaccinum avian influenzae inactivatum immune chicken from infected chicken. The invention also relates to a differential diagnosis reagent kit and its application.

A nano-capsule construct for imaging and therapeutic uses and method for production are provided. One nano-probe embodiment based on genome-depleted plant brome mosaic virus (BMV) whose interior is doped with indocyanine green (ICG), an FDA-approved near infrared fluorescent dye, is used to illustrate the invention. The material encapsulated in viral shell components may be coated with functionalized coatings such as branched, dendritic polymer coatings to improve longevity and distribution in the body as well as antibody conjugation for increased target specificity. The constructs can also be coated with ferromagnetic iron oxide nanoparticles, enabling the ICG-containing capsules to be used as nano-probes with the capability of being detected in both optical and magnetic resonance imaging. The capsules may be produced by purifying a plant or animal viruses and disassembling the viruses to provide virus shell components. The virus shell components are reassembled in the presence of a material for encapsulation thereby encapsulating said material within the core of the construct in one embodiment.

The production process of composite immune globulin for animal includes the following steps: preparing immune blood serum and packing immune blood serum at -20 deg.c inside storing container; separating immune blood serum to extract immune globulin IgG, IgA and IgM; purifying immune globulin IgG, IgA and IgM; mixing immune globulin IgG, IgA and IgM in the ratio of 8-10 to 1 to 1-2; and nanofiltering. The composite immune globulin thus produced includes three kinds of immune globulin, IgG, IgA and IgM, and has three kinds of immune functions, high immune effect of preventing animal viral diseases and easy absorption.

The invention relates to the field of animal virus antibody diagnosis detection, and in particular provides a porcine reproductive and respiratory syndrome virus (PRRSV) antibody detection method and application thereof. The method comprises the following steps: firstly, cloning a non-structure protein NSP4 of a PRRSV HUN4 vaccine strain into a pET-28a (+) prokaryotic expression vector, performing low-temperature induction soluble expression, purifying by utilizing an His marker, and by taking purified protein as coating antigen, establishing an indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) method; confirming a critical value as 0.406, detecting 1274 cases of clinical pig serum by utilizing the ELISA method, detecting by utilizing an American IDEXX kit at the same time, and comparing results, wherein the positive rate of the 1274 cases of clinical pig serum is 71.59%, and the positive rate agreement rate is up to 92%. The detection method provided by the invention is capable of effectively implementing deficiency of the IDEXX kit and can be used as a common PRRSV antibody detection method.

The invention discloses an mRT-PCR (multiple reverse transcription-polymerase chain reaction) kit for discriminating and detecting important avian economic animal viruses and an application of the mRT-PCR kit. The kit comprises primer pairs for detecting an AIV (avian influenza virus) M gene, AIV H7,H9 and H5 subtypes, an NDV (newcastle disease virus), an ILTV (infectious laryngotracheitis virus), an IBDV (infectious bursal disease virus) and an EDS (egg drop syndrome) virus. The mRT-PCR kit has the characteristics of high sensitivity, high specificity and the like when used for detecting AIVs and other common respiratory viruses, can be used for immediately detecting and discriminating important and popular subtypes of the AIVs and five avian respiratory pathogens most popular in China and has a great significance in zoonosis control, monitoring, prevention and control of avian mixed infection, biosafety of farms, reduction of loss of the breeding industry, food safety, quarantine and the like.