207 results about "Porcine parvovirus" patented technology

The present invention provides methods for detecting viral infectivity and content in an enzyme preparation. In certain embodiments, the invention relates to methods for producing a pharmaceutical pancreatic enzyme composition. In additional embodiments, the invention relates to detecting infectious porcine parvovirus (PPV) and determining PPV content in pancreatic enzyme preparations (PEPs), including pancrelipase preparations.

The invention provides a Taqman-MGB fluorescent quantitative PCR kit and a method for detecting 12 common viruses and bacteria of pigs at the same time. The kit comprises PCR reaction liquids A/B/C, wherein the PCR liquids comprise primer pairs and Taqman probes for porcine parvovirus (PPV), type-II streptococcus suis (SS-II), a porcine pseudorabies virus (PRV), type-II porcine circovirus (PCV-2), a hog cholera virus (CSFV), a pig foot and mouth disease virus (FMDV), a porcine reproductive and respiratory syndrome virus (PRRSV), a high pathogenicity porcine reproductive and respiratory syndrome virus strain (Hp-PRRSV), a transmissible gastroenteritis virus (TGEV), an epidemic diarrhea virus (PEDV), rotavirus (PRTV) and a swine influenza virus (SIV) respectively. 12 pathogens of pigs can be detected rapidly and effectively at the same time, the detection method is high in accuracy, specificity and sensitivity and is good in stability, and rapid diagnosis and effective detection on pathogens to be detected can be achieved.

The purpose of the invention is to disclose a porcine circovirus, porcine parvovirus and porcine reproductive and respiratory syndrome virus triplex virus-like particle vaccine and its preparation method. The triple virus-like particle vaccine (Triple VLP vaccine) of the invention contains VLP which is composed of PCV-2 major structural protein CAP protein, PPV VP2 protein epitope and PRRSV Gp5 protein epitope. It is proved by experiment that the vaccine can stimulate good double cellular and humoral immune response. It is shown by pharmacodynamic test that after immunization of different animal groups, the vaccine of injection, nose drops and water forms prepared by VLP antigen formed by the method with or without adjuvants can safely and effectively prevent the infection of PCV-2, PPV and PRRSV. The invention provides an ideal vaccine for the security of sows, piglets and fattening pigs to effectively prevent mixed infection of PCV-2, PPV and PRRSV.

The invention discloses a colloidal gold immunochromatographic test strip for detecting wild-type classical swine fever virus, which consists of water absorbent paper (1), a cellulose nitrate membrane (2), a colloidal gold pad (3), a sample pad (4) and a support (5), wherein the cellulose nitrate membrane contains a detection line which is formed by coating monoclonal antibody HQ06 of anti-classical swine fever virus E2 protein and a quality control line which is formed by coating rabbit anti-mouse IgG antibody; and the colloidal gold pad is combined with colloidal gold-labeled monoclonal antibody 6E10 of the anti-classical swine fever virus E2 protein. The test strip does not react with C-strain of classical swine fever virus, bovine viral diarrhea virus, porcine reproductive and respiratory syndrome virus, transmissible gastroenteritis virus, porcine epidemic diarrhea virus, porcine rotavirus, pseudorabies virus, porcine parvovirus and porcine circovirus type 2, and can accurately and sensitively identify the wild-type classical swine fever virus, thereby having good specificity, sensitivity and repeatability.

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 a primer, probe and assay kit used for detecting porcine circovirus, porcine pseudorabies virus and porcine parvovirus. The primer and probe used for detecting are shown as sequences from SEQIDNO:1 to SEQIDNO:9 in a sequence table. The invention also provides an assay kit used for detecting the porcine circovirus, the porcine pseudorabies virus and porcine parvovirus. The primer and the probe which are adopted by the invention have strong specificity, and three viruses can be detected in one step, thus cost is saved, steps are reduced, and efficiency is improved. The method provided by the invention has the characteristics of high specificity, high sensitivity, high efficiency and low cost and is applicable to analysis of a larger number of samples.

The invention discloses a multiplex RT-PCR detection primer for a porcine delta coronavirus, a porcine epidemic diarrhea virus and a porcine transmissible gastroenteritis virus. The minimum detection capacity of the multiplex RT-PCR for the three viruses is 4.05*10<1> copies/microliter, 4.52*10<3> copies/microliter and 5.47*10<3> copies/microliter respectively. The amplification results for a porcine parvovirus (PPV) and a porcine pseudorabies virus (PRV) are both negative. The multiplex RT-PCR detection results of 57 clinical samples show that one sample is infected with the three viruses at the same time, 11 samples are infected with the PDCoV, 15 samples are infected with the PEDV, one sample is infected with the TGEV, five samples are infected with the PDCoV and the PEDV, and one sample is infected with the PDCoV and the TGEV.

The invention discloses a feed additive for preventing porcine parvovirus infection. The feed additive is prepared from the following bulk pharmaceutical chemicals in parts by weight: 6-8 parts of rhizoma cimicifugae, 4-7 parts of radix scutellariae, 6-8 parts of radix sophorae flavescentis, 6-8 parts of Chinese pulsatilla root, 3-6 parts of java brucea fruit, 3-6 parts of antifeverile dichroa root, 6-8 parts of artemisinin, 6-8 parts of radix stemonae, 4-7 parts of radix isatidis, 4-7 parts of pinellia ternate, 4-7 parts of herba houttuyniae, 3-6 parts of liquorice, 6-8 parts of rhizoma coptidis, 3-6 parts of ash bark and 6-8 parts of radix astragali. The preparation method is characterized by mixing the bulk pharmaceutical chemicals and then grinding the mixture or grinding the bulk pharmaceutical chemicals respectively and then mixing the powder. Through applications, the feed additive has the advantages of good prevention and treatment effects, no toxic or side effect, easily obtained raw materials and low price.

The invention provides a pig virus gene chip and a detection method thereof. The gene chip comprises a probe fixedly arranged on a substrate carrier, wherein the probe is selected from the characteristic segments of the following viruses: a classical swine fever virus (CSFV), a porcine reproductive and respiratory comprehensive virus (PRRSV), a pseudorabies virus (PRV), a porcine circovirus (PCV) and a porcine parvovirus (PPV). The types of the viruses detected by the method are various and the viruses cover common porcine viruses substantially. Random primer polymerase chain reaction (PCR) and multiplex-PCR are adopted to mark, so that false positive which is easy to cause when a PCR result is detected by gel electrophoresis is avoided, and high-flux accurate detection with short time is realized. The substrate carrier is a glass sheet which is subjected to aldehyde treatment, so that the combination between the probe and a target is facilitated, and higher noise is not brought to detection.

The invention discloses a preparation method of inactivated porcine parvovirus vaccine, comprising the following steps: (1) inoculating ST (swine testicle) cells with lentogen strains of porcine parvovirus, culturing to obtain production virus seeds; (2) adding treated microcarriers to a cell growth medium in a bioreactor, stirring; (3) inoculating the ST cells into the microcarriers in the bioreactor for cell reproduction and culturing; (4) infecting the suspension cultured ST cells in the bioreactor with the production virus seeds of the porcine parvovirus for virus reproduction and culturing; and (5) gathering the reproduced virus liquid, inactivating and preparing the vaccine. The method provided by the invention has long cell hold time, high cell viability and high density of living cells, and is favorable for continuous reproduction of virus, so that the prepared virus liquid has high virus titer. The production processes of the method disclosed by the invention are tightly and smoothly linked, the production scale is easy to enlarge, the porcine parvovirus is convenient to gather and stable in quality, and the method can obviously lower the production cost and effectively improve the yield and quality of the vaccine.

The invention discloses a dual SYBR Green I real-time fluorescence polymerase chain reaction (PCR) detection primer and a dual SYBR Green I real-time fluorescence PCR detection method for porcine pseudorabies virus and porcine circovirus type 2. The primer is designed and synthesized, and the dual SYBR Green I real-time fluorescence PCR detection method for the porcine pseudorabies virus and the porcine circovirus type 2 by using the primer comprises the following steps of: extracting deoxyribose nucleic acid (DNA) of a sample; and detecting the sample by using an SYBR Green I real-time fluorescence PCR reaction system and an SYBR Green I real-time fluorescence PCR amplification program. By the primer and the method, two viruses, namely the porcine pseudorabies virus and the porcine circovirus type 2 can be detected simultaneously, and porcine parvovirus, classical swine fever virus, porcine reproductive and respiratory syndrome virus and swine influenza virus cannot be detected. The primer and the method have the characteristics of higher sensitivity, repeatability and stability, and contribute to the identification and the diagnosis of pregnant sow reproductive disturbance virus disease.

The invention discloses porcine parvovirus nanometer alumina gel adjuvant inactivated vaccine which is prepared by mixing inactivated porcine parvovirus virus solution and nanometer alumina gel adjuvant according to the ratio of 1 to 2. According to the invention, safety evaluation to the vaccine is performed on suckling mice and pigs, the results show that after immunization, one suckling mouse dies, no sensitization response is caused on a baby pig, as the same, no obvious full body or partial reaction can be observed, so that the inactivated vaccine is fully proved to be safe and reliable. The porcine parvovirus nanometer alumina gel adjuvant inactivated vaccine set has a CD4 +/CD 8 + relative value in the fourth immunization week, which is obviously higher than that of other vaccine set, the relative value is increased to 2.97 from 2.11, which shows that the vaccine enhances the immune state of cells of mice. The porcine parvovirus nanometer alumina gel adjuvant inactivated vaccine provided by the invention can induce the secretory expression of IFN-Gamma, and the vaccine can remarkably enhance the immune response reaction produced by Th 1 type cells, to induce the Th 1 type cells to secrete IFN- Gamma. Therefore, the immune level of cells can be increased, and a stronger cellular immunity response can be produced.

The invention provides a special kit for detecting highly pathogenic porcine reproductive and syndrome virus mutation strain. The highly pathogenic porcine reproductive and syndrome virus mutation strain is detected by self-designing specific primers PSX1/PSX2, extracting virus RNA and carrying out reverse transcription into cDNA and utilizing a PCR method; the size of the estimated amplification segment of an amplification mutation strain is 404bp and an amplification traditional PRRSV amplification segment is 494bp. Compared with the prior art, the special kit has strong specificity and sensitivity, and the coincidence rate can achieve more than 90 percent compared with a virus separation and IPMA method. The amplification results of the primers to hogcholera virus, pseudorabies virus and porcine parvovirus are negative, which can distinguish a NSP2 mutation strain and a traditional strain; the method can detect the PRRSV with 12.8ng/L, thus the invention can be widely used for the clinical detection of the highly pathogenic porcine reproductive and syndrome virus mutation strain.

The invention discloses a porcine parvovirus L strain and use thereof in preparation of porcine parvovirus inactivated vaccines. In the invention, microbial preservation number of the separated strain is CGMCC No. 3352. The strain is highly homologous with an NADL-2 strain and a China strain and keeps high reproduction, and the viruses are stable. Oil adjuvant is added into antigen liquid which inactivates the viruses by using binary ethylenimine to prepare the bidirectional oil-emulsion inactivated vaccines; and the prepared inactivated vaccines are applied in a vaccine inoculation experiment of 10,276 pigs (most of which are first farrowing sows), and in the experiment, an immune dosage of 2 millimeters is given to each pig through muscle injection, and each time of inactivated vaccine inoculation can obtain an immune period of over 6 months. Under the conditions that a 10 millimeter overdose vaccine is injected into a pig and that single dose inoculation is repeatedly carried out for three times, no abnormal reaction happens, which proves that the inactivated vaccines have high immunogenicity and safety.

The invention discloses a porcine parvovirus assay kit and application thereof. The kit provided by the invention comprises three pairs of primers, namely an inside primer pair, an outside primer pair and an annular primer pair combined with an NS1 gene in genome DNA (GenBank Accession Number NC-001718) of a porcine parvovirus. The porcine parvovirus assay kit of the invention has the advantage of high assay sensitivity and is simple and convenient for operation, can detect ten copied target DNAs at minimum and is particularly suitable for clinical medicinal detection and the detection of porcine parvovirus probably polluting foods in the basic level yield.

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 provides porcine parvovirus, vaccine composition and application thereof, and belongs to the technical field of biology. The microbial preservation number of the porcine parvovirus PPV-JS strain is CGMCC (China General Microbiological Culture Collection Center) No.6605. The vaccine composition provided by the invention comprises an inactivated porcine parvovirus PPV-JS strain and veterinary pharmaceutically acceptable adjuvant. The invention also provides the application of the porcine parvovirus PPV-JS strain in preparing medicaments for preventing diseases caused by porcine parvovirus. The porcine parvovirus PPV-JS strain has excellent immunogenicity, and can be used as an activated vaccine production strain and a virus seed for inspection. The vaccine composition (porcine parvovirus inactivated vaccine) provided by the invention is good in safety, high in immune efficacy and long in immunity period, and only requires single dose, so that manpower and side effect of the vaccine to a target animal can be greatly reduced.

The invention relates to a method for producing a porcine parvovirus inactivated vaccine by using a torrent bioreactor, comprising the following steps: inoculating the porcine parvovirus into the IBRS-2 cell, and culturing to obtain a production virus strain; inoculating the IBRS-2 cell into the torrent bioreactor; inoculating the prepared porcine parvovirus strain into the IBRS-2 cell cultured in the torrent bioreactor, and proliferating and culturing the virus; harvesting the proliferated virus solution; inactivating; and matching to obtain the porcine parvovirus inactivated vaccine. According to the method, because the torrent bioreactor and a paper vector are used, the shearing force generated by a microcarrier bioreactor used for culturing the cell is avoided, the density and the viability of the cell are further improved, and the capacity of proliferating the viruses by the cell is enhanced. The reactor can be used for monitoring and automatically regulating the optimal condition for growing the cells, thus the viruses harvested in all bathes have small difference, the quality stability of the vaccine is improved, the vaccine production efficiency is improved, and the vaccine production cost is reduced.

The invention relates to polyvalent vaccines for prevention and treatment of porcine infectious diseases, especially to a combined vaccine for treatment and prevention of porcine circovirus type 2 (PCV2) and porcine parvovirus. By selecting PCV2 and porcine parvovirus, the preparation method provided in the invention consists of: culturing PCV2, and conducting inactivation and concentration; culturing a porcine parvovirus, and performing inactivation and concentration; mixing the two antigenic components in proportion, and supplement an adjuvant to prepare the vaccine. The bivalent vaccine prepared in the invention is easy to use, more secure, and has an immune effect superior to that of combined use of two single vaccines.