205 results about "Porcine Circoviruses" patented technology

An improved method for recovering the protein expressed by open reading frame 2 from porcine circovirus type 2 is provided. The method generally involves the steps of transfecting recombinant virus containing open reading frame 2 coding sequences into cells contained in growth media, causing the virus to express open reading frame 2, and recovering the expressed protein in the supernate. This recovery should take place beginning approximately 5 days after infection of the cells in order to permit sufficient quantities of recombinant protein to be expressed and secreted from the cell into the growth media. Such methods avoid costly and time consuming extraction procedures required to separate and recover the recombinant protein from within the cells.

The present invention belong to veterinary new biological medicine technology field, relates to porcine circovirus type II (PCV2) inactivated vaccine of and method for preparing same. The vaccine used seed virus is porcine circovirus type II DBN-SX07 strain, the preservation number is CGMCC No 3064, the virus strain is used as antigen preparation of inactivation by alkyl agents and emulsification by adding oil adjuvant. Using the invention provided PCV2 inactivated vaccine to immune pig can generate an uniform and effective protection force-shorting PCV2 infection windows period obviously, and prolong immune duration-reducing times of booster immunization.

This invention provides a trivalent immunogenic composition including a soluble portion of a Mycoplasma hyopneumoniae (M. hyo) whole cell preparation; a porcine circovirus type 2 (PCV2) antigen; and a PRRS virus antigen, wherein the soluble portion of the M. hyo preparation is substantially free of both (i) IgG and (ii) immunocomplexes comprised of antigen bound to immunoglobulin.

The invention relates to construction and application of recombinant nucleocapsids of Cap genes of porcine circovirus expression type 2 nucleocapsid protein, belonging to the genetic engineering bacterin field. C-terminal gene fragments of porcine parvovirus (PPV) VP2 genes are cloned into a type 5 adenovirus shuttle vector of the human beings, and recombinant adenovirus rAd-deltaVP2 is obtained; deltaVP2 proteins are expressed successfully and highly efficiently and can be self-assembled into the nucleocapsids [PPV:VLPs]; the PPV VP2 nucleocapsids are used as antigen transport vectors and 165 to 200 sites of amino acid (deltaCap) genes of the porcine circovirus type 2 (PCV2) nucleocapsid proteins (Cap) are embedded into an N-terminal (deltaVP2) of the PPV VP2, and then recombinant adenovirus rAd-deltaCap-deltaVP2 is obtained; embedded VP2 (deltaCap-deltaVP2) proteins are expressed successfully and highly efficiently and can be self-assembled into nucleocapsids [PPV:VLP(PCV2)]. The invention also relates to application of the recombinant virus and recombinant PPV VP2 nucleocapsids of the expression Cap genes of the recombinant virus in the aspects of bacterin immunity and so on.

The invention discloses an RPA (recombinase polymerase amplification) primer and a detection kit for rapidly detecting African swine fever viruses. Target genes can be effectively amplified, specificity is 100%, detection sensitivity is 102 copy/reaction, and sensitivity is equivalent to that of fluorescent quantitative PCR (polymerase chain reaction). Cross reaction between the RPA amplificationprimer and one of classical swine fever viruses, vesicular exanthema swine viruses I, porcine reproductive and respiratory syndrome viruses, porcine circoviruses and the like is omitted. A RPA isothermal amplification system is rapidness in reaction and wide in temperature range, effective amplification of the target genes can be achieved at the temperature of 38-46 DEG C, and the detection kit can rapidly, efficiently and sensitively detect the African swine fever viruses and has the advantages that the kit is simple to operate, high in specificity, safe and free from pollution, reaction results are easily observed and the like. Effective technical means are provided for on-site rapidness detecting and screening of infection nucleic acid of the African swine fever viruses, and the RPA amplification primer has great significance for control of infection spreading of the African swine fever viruses in China and inspection and quarantine in infected areas and entry and exit ports.

The invention provides a porcine circovirus 2d-type and 3-type Cap protein bivalent subunit vaccine and a preparation method and application thereof. By cloning porcine circovirus 2d-type and 3-type Cap protein epitope genes, an insect-baculovirus expression vector system is adopted to successfully express and obtain high-purity 2d-type and 3-type Cap proteins. The porcine circovirus 2d-type and 3-type Cap proteins bivalent subunit vaccine is developed for the first time by utilizing the two expressed proteins. The prepared bivalent vaccine is strong in immunity and high in safety, can achievean ideal immune protection effect on the 2d type and 3 type porcine circoviruses, and provides an effective means for prevention and control of the porcine circoviruses.

The invention relates to a method for preparing a genetic engineering product, in particular to a prokaryotic expression protein of VP73 gene from African swine fever virus (ASFV) and a preparation method thereof. The preparation method comprises: artificially synthesizing the whole-length sequence of VP73 gene according to the sequence of the VP73 gene from ASFV in GenBank, constructing a recombinant expression vector pET32a-VP73, sequencing, verifying, transforming prokaryotic expression recipient bacteria E.coli BL21(DE3), and inducing expression by isopropyl-1-thio-beta-d-galactopyranoside (IPTG), wherein the molecular weight of the recombinant fusion protein is about 65KD. Protein purified by nickel column affinity chromatography can undergo a specific immune imprinting reaction with ASFV positive serum and avoid cross reaction with viruses such as swine fever virus, hog cholera virus, porcine circovirus, porcine reproductive and respiratory syndrome virus, swine influenza virus and pseudorabiesvirus. Experiments show the expressed protein has high detection sensitivity, and high specificity. When the antigen is used for detection, risk of spreading poison is avoided. And the antigen can be used as a detection antigen for use in an enzyme-linked immuno sorbent assay (ELISA) method for identifying an ASFV antibody.

The invention discloses an ELISA (enzyme linked immunosorbent assay) kit for porcine circovirus type 2 (PVC2) antibody detection. The kit includes an antibody detection board, a sample diluting solution, an enzyme-labeled antigen, a washing solution, a substrate color developing solution, a stop solution, a positive serum and a negative serum. The Cap protein coated with antibody detection board of the kit is PCV2 eukaryotic expression nucleocapsid Cap protein, and the non-specific reactions are greatly reduced by using the Cap protein coupled enzyme-labeled antigen, so that the antibody detection accuracy is improved and the kit is more favorable for large-scale popularization and use.

The invention provides a vaccine composition comprising an immune amount of a porcine circovirus type 2 antigen, an immune amount of porcine reproductive and respiratory syndrome virus antigen, a porcine parvovirus antigen and a carrier acceptable in veterinary medicine. Antigens in the vaccine composition have no mutual interference on each other, even show some immune synergism; the composition causes small stress after immunization, is more safe, has immune effect better than that of a single vaccine, avoids the stress response caused by multiple inoculation, simplifies the complex immune procedure, and reduces the cost of epidemic prevention.

The invention discloses a traditional Chinese medicine composition for treating porcine circovirus disease and a preparation method thereof. The traditional Chinese medicine composition comprises the following traditional Chinese medicines: astragalus root, iastis root, forsythia fruit, honeysuckle, cordyceps polysaccharide, codonopsitis, atractylodes rhizome, angelica, fresh rehmannia root, Chinese goldthread, grifola umbrellata, tuckahoe, white atractylodes rhizome, tangerine peel, and epimedium. The preparation method comprises the following steps: 1) respectively attritioning and screening various components except the cordyceps polysaccharide; 2) uniformly mixing the astragalus root, codonopsitis, forsythia fruit, atractylodes rhizome, angelica, grifola umbellate, tuckahoe with the cordyceps polysaccharide; 3) uniformly mixing the iastis root, honeysuckle, fresh rehmannia root, Chinese goldthread, white atractylodes rhizome, tangerine peel, and the epimedium; 4) uniformly mixing the medicine power obtained from the step 2) and the step 3), so as to obtain the traditional Chinese medicine composition. The traditional Chinese medicine composition is simply prepared, safe and reliable, and has no toxic and side effect, high bioavailability, and reliably and effectively prevents and treats the porcine circovirus disease, and moreover promotes the growth of the pit and is widely used in clinical.

The present invention relates to the use of an immunogenic composition comprising a porcine circovirus type 2 (PCV2) antigen for the prevention and treatment of sub-clinical PCV2 infection in animals, preferably in pigs.

The present invention provides for pharmaceutical compositions comprising pancreatic enzyme preparations (PEPs) with viral infectivity reduced below significant levels and having high enzymatic activity. The PEPs can comprise lipases, proteases, amylases, non-enveloped viruses (e.g., porcine parvovirus (PPV), porcine circovirus type 2 (PCV-2), porcine encephalomyocarditis virus (EMCV)), and enveloped viruses (e.g., vesicular stomatitis virus (VSV), and influenza A (IFA)). The present invention also includes methods of treating pancreatic insufficiency by administering these pharmaceutical compositions and methods of making the same by treating the PEP with beta-propiolactone (BPL) to reduce viral infectivity.

The invention discloses a method for synchronously detecting a porcine circovirus type 2 (PCV2), a classical swine fever virus (CSFV) and a porcine reproductive and respiratory syndrome virus (PRRSV). The method comprises the following steps of: designing three pairs of specific probes by the PRRSV nucleic acid, PCV2 nucleic acid and CSFV nucleic acid, preparing reporter genes of probe markers by using polymerase chain reaction (PCR), and implementing loop-mediated indirect PCR detection of three viruses by cyclization and reverse PCR amplification of the reporter genes. the pathogens of PRRSV, PCV2 and CSFV in sample tissues can be synchronously detected by designing three pairs of specific probes aiming at three viruses, marking the same reporter genes, designing a pair of reverse PCR amplification primers and establishing a loop-mediated indirect PCR detection system, so that certain reference is provided for scientifically preventing and controlling porcine diseases. Clinical tests show that the method has the advantages of single detection background, high sensitivity, strong specificity, good stability and the like, can be used for single-virus detection, can also be used for synchronous detection of multiple viruses, and provides a tool for viral clinical diagnosis and epidemiological study of PRRSV, PCV2 and CSFV.

The invention discloses a multiplex PCR detection kit for identifying porcine circovirus (PCV). The kit comprises the following three pairs of multiplex PCR detection primers which can quickly distinguish genotype PCV1, genotype PCV2 and genotype PCV3: PCV1-F and PCV1-R, PCV2-F and PCV2-R, and PCV3-F and PCV3-R.The multiplex PCR detection kit disclosed by the invention can be used for detecting the PCV and distinguishing different genotypes, has the characteristics of being quick, simple, convenient, high in specificity and high in sensibility, and can perform large batches of sample detectionat the same time; only one strand displacement amplification is needed, so that the situation whether a sample is infected by PCV1, PCV2 or PCV3, or is jointly infected by two or three of the PCV1, the PCV2 and the PCV3 can be identified; and the multiplex PCR detection kit can provide technical support for epidemic situation surveillance, epidemiologic investigation and comprehensive preventingand controlling of the PCV, and has a favorable application prospect.

The invention belongs to the technical field of animal virology and molecular biology and discloses a duplex PCR (polymerase chain reaction) detection primer and kit for quickly distinguishing porcinecircoviruses type 2 and type 3. The kit comprises primer sequences shown as SEQ ID1-4, 2*F8 Fastlong PCR MasterMix, a cDNA template and sterile water. The duplex PCR detection kit for quickly distinguishing the porcine circoviruses type 2 and type 3 has advantages that by duplex PCR amplification, two types of DNA viruses similar in lesion can be detected in one time, the total detection time iscontrolled to be about two hours, a detection method is easy in operation, convenient and quick, and quick detection can be realized.