24 results about "Circovirus" patented technology

The present application belongs to the field of molecular biology detection, and particularly relates to a composition, method and use for simultaneous detection and differentiation of swine fever virus, psittacosis chlamydia, porcine reproductive and respiratory syndrome virus and porcine circovirus type II. The composition for simultaneous detection provided by the present application can realize rapid detection of four pathogen target genes in one tube of reaction solution by using multiplex fluorescence RPA detection technology, and has the advantages of high throughput, short time and low cost. On one hand, the present application can accurately detect pathogens and analyze mixed infection of pathogens, so as to achieve the effect of precise medication, reduce the use of antibiotics, save costs, and reduce the cost and increase the benefit of enterprises, and on the other hand, compared with fluorescence PCR, the detection time is greatly shortened, and the purpose of rapid detection is achieved.

The present application relates to the technical field of protein purification, and particularly relates to a porcine circovirus vaccine protein purification device, which comprises a protein purification instrument, a tray is arranged on the top of the protein purification instrument, and a connecting mechanism is arranged at the connecting position of the tray and the protein purification instrument. The tray is installed and fixed through the mutual insertion of the convex block and the pin rod, so that the tray is more convenient to take down and install and is easy to clean. The internal inclined surface of the tray achieves the effect of flow guiding, liquid is stored in the water collecting box, dripping on the experiment table is avoided, the safety of the experiment is improved, the height adjustment of the bracket is more convenient through the cooperation of the arc-shaped clamping block and the arc-shaped groove, the positioning effect is achieved through the cooperation of the spherical convex and the spherical groove, the adjustment of the installation position of the pre-packed column is more convenient, and the operation is more simple and convenient during the protein purification operation.

The invention discloses an mRNA vaccine composition for combined treatment of porcine circovirus and application of the mRNA vaccine composition. The combination comprises a nucleotide combination, the nucleotide combination comprises a sequence for coding the Cap protein of the porcine circovirus and a CpG adjuvant, and the sequence for coding the Cap protein of the porcine circovirus and the CpG adjuvant are respectively assembled into a porcine circovirus mRNA vaccine and a CpG adjuvant mRNA vaccine. In order to solve the problem that the existing mRNA vaccine generally depends on inherent immune stimulation of mRNA and LNP, and generally, an exogenous adjuvant is not additionally added, so that the immune response intensity or persistence is insufficient possibly, the CpG adjuvant and the PCV2 Cap mRNA-LNP vaccine are creatively combined for use; experiments show that the combination can significantly improve the neutralizing antibody level and enhance the immune response of Th1 type cells compared with mRNA independent immunization, has good safety, and can be used for preventing and / or treating PCV2 infection.

The invention provides a porcine circovirus type 5 Cap protein, a virus-like particle, an elisa plate, an ELISA kit and application thereof, and belongs to the technical field of animal viruses. The invention provides a porcine circovirus type 5 Cap protein with an amino acid sequence as shown in SEQ ID NO. 1. The porcine circovirus type 5 Cap protein provided by the invention has the advantages of stable structure and good antigenicity, can be specifically combined with a circovirus type 5 antibody in a serum sample, and can be self-assembled into 60 virus-like particles. The ELISA plate and the ELISA kit for detecting the porcine circovirus type 5, provided by the invention, have the advantages of strong specificity and high sensitivity, and can accurately detect whether a blood sample contains the porcine circovirus type 5 or not. The porcine circovirus type 5 virus-like particle provided by the invention has the advantages of high yield, good uniformity and strong stability, and can be subsequently prepared into a porcine circovirus type 5 subunit vaccine.

The application belongs to the technical field of biotechnology, and particularly discloses a synthetic peptide antigen of porcine circovirus type 2 Cap protein, a polyclonal antibody and application thereof. In order to solve the technical defects of low specificity and inaccurate detection of the amino acid sequence of the synthetic peptide antigen in the prior art, the application provides a synthetic peptide antigen of PCV2 Cap protein, a polyclonal antibody and application thereof. The application obtains an amino acid sequence of an antigen peptide by performing antigen peptide prediction on PCV2 Cap protein, and the applicant synthesizes the antigen peptide by using an Fmoc solid phase method, and immunizes a New Zealand white rabbit with the obtained antigen peptide to prepare a polyclonal antibody of Cap protein, the antibody can effectively react with a transiently expressed Cap protein and a Cap protein in PCV2b-infected PK15 cells, and provides a powerful tool for detection and biological function research of the PCV2 Cap protein.

This invention relates to the field of cell culture technology, specifically to a cell culture tank for producing porcine circovirus vaccine. The tank includes a main body with a feeding port at the top, a fixing box at the top, and a heat dissipation vent at the top of the fixing box. A stirring mechanism is located inside the main body. The first cone design achieves a shearing and flow-diverting effect, the arc surface guides the flow, reducing cell damage, and the spherical protrusions and through-holes further enhance the flow disturbance effect. The row of staggered spherical protrusions further improves the mixing effect. The two flow-dispersing blocks are arranged in opposite directions, achieving mixing in both forward and reverse rotation. The second cone design reduces resistance during reverse rotation without affecting mixing.

The present invention in particular relates to a method for reducing virus shedding, colonization of lymphoid tissue, and / or lymphoid depletion caused by or associated with a porcine circovirus type 2 (PCV2) infection in a pig, the method comprising administering orally to the pig an immunogenic composition comprising non-replicating PCV2 antigen. In one example, an immunogenic composition is administered via the oral route to a pig, wherein the immunogenic composition comprises a recombinantly produced and purified PCV2 ORF2 protein. This allows to simply reduce virus shedding, colonization of lymphoid tissue, and / or lymphoid depletion caused by associated with a PCV2 infection in the pigs, while simultaneously reducing the propensity for adverse reactions, eliminating the risk of injection site reactions, avoiding pain, and putting less stress on the animals.

The application discloses a bovine viral diarrhea virus chimeric virus expressing porcine circovirus type 3 capsid protein and a preparation method thereof. In the experiment, a gene sequence for encoding a porcine circovirus type 3 capsid protein and a His tag fused with the C terminal of the protein is inserted into different sites of a bovine viral diarrhea virus genome through a circular polymerase extension reaction, and the chimeric virus is prepared through virus rescue of the obtained circular DNA. The His tag sequence in the gene sequence and the bovine viral diarrhea virus genome are connected through a 2A peptide sequence, so as to realize the target of effectively releasing the antigen through the co-expression of the heterologous protein. The chimeric virus prepared by the application retains the infectivity and replication capacity of the parent virus, can stably replicate in cells and efficiently express the target antigen, and provides an efficient and flexible virus carrier platform for the research and development of a porcine circovirus type 3 vaccine.

The application belongs to the field of biological detection, and particularly relates to an isothermal amplification kit for detecting porcine circovirus type 3. The application provides an isothermal amplification detection primer group for porcine circovirus type 3 (PCV3), which comprises a pair of outer primers (LOF and LOR) and a pair of inner primers (LIF and LIR) and is designed according to the conserved sequence of the porcine circovirus type 3. The primer group has low design difficulty, and maintains similar sensitivity and specificity to other isothermal amplification reactions such as a loop-mediated isothermal amplification method, can be suitable for target sequences with higher GC content and below 200 bp, has no denaturation and annealing process, is suitable for on-site detection, and has great application potential.

The invention provides a subunit vaccine composition for swine fever, porcine pseudorabies, porcine circovirus and swine erysipelas as well as a preparation method and application of the subunit vaccine composition. The subunit vaccine composition comprises prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein, viral subunit protein and pharmaceutically acceptable adjuvants. Wherein the amino acid sequence of the prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein is as shown in SEQ No.1, and the viral subunit protein is selected from one or more of subunit E2 protein of hog cholera virus, gD protein of porcine pseudorabies virus, gB protein of porcine pseudorabies virus and cap protein of porcine circovirus type 2. The vaccine composition disclosed by the invention has the advantages of strong immunogenicity, good safety, no immune interference, high neutralizing antibody titer and long antibody duration; the infection of swine erysipelas filamentous bacillus can be effectively prevented and protected.

This invention discloses a porcine circovirus vaccine filling device, relating to the field of vaccine filling technology. The device includes a filling assembly comprising a base, a support frame on top of the base, the support frame being L-shaped, a servo motor at the top of the support frame, and a connecting frame fixedly mounted on the bottom telescopic end of the servo motor through the support frame. The connecting frame has several sets of mounting holes arranged sequentially, through which connecting pipes are installed. This invention, by setting up the filling assembly, can effectively buffer the filling process. The sieve plate provides effective buffering and defoaming, solving the problem of air bubbles easily generated during filling in existing devices. It avoids the problem of large differences in drug dosage due to air bubbles, ensuring the product complies with veterinary pharmacopoeia regulations. Simultaneously, it prevents oxygen in the air bubbles from accelerating the oxidative denaturation of vaccine antigen proteins, ensuring the drug's efficacy and preventing immunization failure.

The invention relates to the technical field of biological medicines, and particularly discloses a vaccine adjuvant composition as well as a preparation method and application thereof. The vaccine immune composition provided by the invention comprises GM-CSF and cortex albiziae saponin, and has the following advantages: 1, the vaccine immune composition can significantly improve the specific antibody level and the cytokine secretion efficiency, effectively promote the activation of killer T cells, efficiently induce the response of memory T cells, and construct a stable immune defense basis; 2, the immune level of the organism can be remarkably improved after challenge, the tissue injury risk is reduced by inhibiting replication and colonization of viruses in key target organs, and meanwhile, the body weight of the organism is promoted to be quickly recovered after challenge, so that the physiological homeostasis of the organism is better maintained; thirdly, the method is wide in adaptability and can be compatible with various veterinary vaccines such as foot-and-mouth disease vaccines, porcine reproductive and respiratory syndrome vaccines, pseudorabies vaccines and porcine circovirus type 2 vaccines. The method can be applied to the fields of research, development and preparation of vaccine adjuvants or vaccines and the like.

The application provides a pig getah virus positive serum and a preparation method and application, and belongs to the technical field of animal virology. The application provides a preparation method of a pig getah virus positive serum, wherein a CDCD pig is used as a host, neutralizing antibody titer obtained through multiple immunization of high-concentration antigen is greater than or equal to 1:512; the serum does not have cytotoxicity affecting cell culture; the CDCD pig is a pig free from maternal antibody interference, which is obtained through caesarean section, isolated feeding and artificial feeding; the positive serum does not detect pig pseudorabies virus, swine fever virus, porcine circovirus type 2, pig foot-and-mouth disease virus, bovine viral diarrhea virus, porcine infectious gastroenteritis virus, porcine epidemic diarrhea virus, porcine rotavirus, porcine parvovirus, porcine Japanese encephalitis virus antibody, and has the characteristics of high titer and no cytotoxicity.

The invention pertains to a combination of a first vaccine comprising anon-replicating immunogen of porcine circovirus type 2 (PCV-2) and a non-replicating immunogen of Mycoplasma hyopneumoniae, and a second vaccine comprising a live attenuated porcine reproductive and respiratory syndrome (PRRS) virus, for use in prophylactically treating a pig against an infection with PCV-2, an infection with Mycoplasma hyopneumoniae and an infection with PRRS virus, by associated separate injection of the first vaccine and the second vaccine into a tissue of the pig at a first and a second injection site respectively, wherein the first and second injection sites are at most 5 cm apart from each other.

The invention belongs to the technical field of livestock epidemic prevention and detection, and provides a donkey circovirus recombinant protein and application thereof. The amino acid sequence of the donkey circovirus recombinant Cap protein is as shown in SEQ ID NO: 1. The donkey circovirus recombinant Cap protein can be used for preparing a reagent for detecting donkey circovirus.

The application relates to the technical field of pathogen detection, and discloses a pathogen multiplex PCR rapid detection method based on GenomeLabTM GeXP multiplex gene expression analysis technology. A primer group composed of seven pairs of specific primers, one pair of positive control primers and one pair of universal primers is used to detect pathogens. The seven pairs of specific primers are respectively used for specifically combining with target genes of porcine fever virus, porcine reproductive and respiratory syndrome virus, pseudorabies virus, porcine parvovirus, porcine circovirus type 3, necrotic fusiform bacteria and porcine epidemic diarrhea virus. The 5' end of the universal upstream primer is provided with a fluorescent label. A two-step PCR amplification program is adopted, and rapid and accurate detection of the seven important pathogens, i.e. porcine fever virus, porcine reproductive and respiratory syndrome virus, pseudorabies virus, porcine parvovirus, porcine circovirus type 3, necrotic fusiform bacteria and porcine epidemic diarrhea virus, is successfully realized in one reaction system.

The application relates to the technical field of biology, and particularly provides an antibody specifically combined with a porcine circovirus type 3 Cap protein and products and applications thereof. The porcine circovirus type 3 Cap protein provided by the application is encoded by a sequence SEQ ID No. 1 or a degenerate sequence thereof, the antibody heavy chain variable region prepared by the application is SEQ ID No. 2, the light chain variable region is SEQ ID No. 4, and the two can be combined to prepare a porcine circovirus type 3 blocking ELISA antibody detection kit, so that the PCV3 infection state can be accurately evaluated.

The invention is applicable to the technical field of veterinary biomedicine, and provides a nano-antibody specifically bound with porcine circovirus type 2 and application thereof, nucleotide sequences of the nano-antibody are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 2, and amino acid sequences of the nano-antibody are respectively shown as SEQ ID NO: 3 and SEQ ID NO: 4. The nano antibody provided by the invention can be specifically combined with porcine circovirus particles, virus neutralizing activity is shown in PK15 cells, and the multivalent nano antibody obtained by using a ferritin coupling system can significantly reduce PCV2 virus particle infection and replication and proliferation capacities.

The invention relates to a device for detecting porcine reproductive and respiratory syndrome virus infectious diseases. Specific primers and fluorescent probes aiming at porcine reproductive and respiratory syndrome virus, porcine parvovirus and porcine circovirus type II are preloaded in a PCR (polymerase chain reaction) tube; the PCR reaction tubes are installed on the detection circular truncated cone in an annular array mode, the detection circular truncated cone is provided with the fluorescence excitation unit, the LED light source emits light with the corresponding wavelength, and the light is irradiated to each PCR reaction tube after being filtered by the optical filters which are arranged in the same annular array mode; all the PCR reaction tubes are driven by respective corresponding driving assemblies to synchronously move along the radial direction of the detection circular table; all the shading cylinders on the mounting disc above the detection circular table can completely cover the PCR reaction tube; a PCR reaction tube is correspondingly arranged on one side between each pair of photosensitive devices at the upper part of the disc edge of the mounting disc; and the photosensitive devices are used for sensing light filtered by the optical filter. The structure is simple and compact, and the detection efficiency and precision can be improved.

The invention provides an antigen polypeptide for preparing a porcine circovirus type 3 (PCV3) synthetic peptide vaccine, a polypeptide composition and the vaccine. The antigen polypeptide is a polypeptide as shown in a sequence 1 or a sequence 2. The polypeptide composition comprises a polypeptide as shown in a sequence 1 and a polypeptide as shown in a sequence 2. The vaccine comprises polypeptide or a polypeptide composition. The PCV3 synthetic peptide vaccine provided by the invention has good immune efficacy and is safe to piglets and pregnant sows. After challenge, PCV3 infection can be effectively treated, and the protection on the body is improved. The vaccine provided by the invention has no biological safety, is easy to synthesize on a large scale, and has a good application prospect. The invention also provides a preparation method and application of the polypeptide and the polypeptide vaccine.

Disclosed are a fusion antigen of porcine Getah virus (GETV), a kit, a preparation method therefor and an application thereof. The fusion antigen of the GETV is primarily prepared by recombining a Gaussia luciferase (GLuc) gene with a codon-optimized GETV E2 antigen gene to construct an expression vector, and transfecting the expression vector containing GLuc-E2 into mammalian cell lines, resulting in the secretion of GLuc-E2 proteins into a cell supernatant for expression. Without the need for protein purification step, the cell supernatant may be directly collected for disease detection. The present disclosure demonstrates strong specificity and shows no cross-reactivity with African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), or Japanese encephalitis virus (JEV).

The invention discloses a recombinant antigen protein, a recombinant vector, a recombinant host cell and application thereof and a vaccine, and relates to the technical field of antigen preparation, the recombinant antigen protein comprises a capsid protein of porcine circovirus type 2 and a polypeptide in a capsid protein of porcine circovirus type 3; the polypeptide in the porcine circovirus type 3 capsid protein comprises B cell neutralization site polypeptide and / or T cell neutralization site polypeptide. The recombinant antigen protein disclosed by the invention can react with PCV2 type positive serum and PCV3 type positive serum at the same time; the vaccine prepared by mixing the recombinant antigen protein with an adjuvant is an efficient bivalent vaccine, and can effectively stimulate a pig body to generate high-concentration specific antibodies of PCV2 and PCV3, so that a reference is provided for joint prevention of the two viruses. The recombinant antigen protein has the characteristics of stable expression, high yield and good immunogenicity.

The invention relates to the technical field of recombinant viruses, and particularly discloses a recombinant porcine Seneca virus for expressing a porcine circovirus 2d genotype Cap gene as well as a preparation method and application of the recombinant porcine Seneca virus. According to the invention, an SVA virus HeN-1 / 2018 is taken as a vector, an infectious cloning vector of a full-length genome of the SVA virus HeN-1 / 2018 is constructed, a PCV2d Cap-P2A gene synthesized by a PCV2d Cap gene without a nuclear localization signal and a P2A gene is recombined into the infectious cloning vector through a seamless cloning technology to obtain a connection product, the connection product is converted into a competent cell, and the recombinant PCV2d Cap-P2A gene with the nuclear localization signal removed is obtained. A recombinant plasmid pSVA-PCV2d-Cap is obtained after culture and verification, then an SVA recombinant virus rSVA-PCV2d-Cap for expressing PCV2d Cap protein is obtained through rescue based on a CMV polymerase transcription system, and the whole gene sequence of the recombinant virus is shown as SEQ ID NO: 1; an inactivated vaccine is prepared from the recombinant virus, it is determined that a vaccine composition produced by the recombinant virus rSVA-PCV2d-Cap strain can induce immune animals to generate neutralizing antibodies with high titers for SVA and PCV2, and a certain theoretical and practical foundation is laid for clinical use of the inactivated vaccine for the SVA and PCV2.

The invention provides a porcine giant circovirus as well as a culture and separation method and application thereof, and belongs to the technical field of biotechnology and veterinary medicine. The invention provides a porcine giant circovirus gene. The nucleotide sequence of the porcine giant circovirus gene is shown as SEQ ID NO. 1. The invention also provides an application of the Marek's disease tumor cell line MSB-1 in separating and culturing the porcine giant circovirus. The porcine giant circovirus is successfully cultured on the Marek's disease tumor cell line MSB-1 for the first time, and high-titer virus liquid enough for vaccine preparation can be obtained. The porcine giant circovirus obtained by separation can be used as a totivirus inactivated vaccine, has the advantage of inducing a high-level antibody after being inactivated and added with an adjuvant to immunize a mouse, and can be subsequently prepared into a porcine circovirus type 5 inactivated vaccine. According to the invention, a key vaccine production platform is directly provided for preventing and controlling diseases such as porcine reproductive disorder caused by the porcine giant circovirus, and the blank of a porcine giant circovirus disease prevention and control technology is filled.