166 results about "Epidemic diarrhea" patented technology

The invention relates to a method for inhibiting in-vitro infection of a porcine epidemic diarrhea virus by using JPH203 and application. The JPH203 can be used for preparing an antiviral drug for inhibiting infection of the porcine epidemic diarrhea virus. JPH203 with the concentration of 5 [mu] M is added in the process that Vero cells are infected with PEDV, compared with a control group without JPH203, the virus infection condition, the protein expression quantity, the RNA content and the virus titer of the PEDV in a treatment group with JPH203 are all remarkably reduced, and it is indicated that JPH203 can be used for inhibiting in-vitro infection of the porcine epidemic diarrhea virus.

The invention relates to the technical field of biological detection, relates to a multiple ddPCR detection method for porcine intestinal coronavirus, and in particular relates to a monitoring and quantitative detection method for samples with relatively low porcine intestinal coronavirus content or complex matrix, such as feeds, animal tissues and animal-derived products. The droplet digital PCR detection method for the porcine intestinal coronavirus established in the invention has the advantages of strong specificity, high sensitivity, good repeatability and strong anti-interference capability, can be used for absolute quantification of the copy concentration of the porcine acute diarrhea syndrome coronavirus (SADS-CoV), the porcine epidemic diarrhea virus (PEDV), the porcine delta coronavirus (PDCoV) and the porcine transmissible gastroenteritis virus (TGEV) in an actual sample, and can be used for the detection of the porcine intestinal coronavirus. The invention provides a novel and reliable technical method for trace early warning and monitoring of early-stage infection of four porcine intestinal coronavirus, differential diagnosis of other common viruses, research of propagation rules of the viruses, epidemiological risk assessment and other systematic applications.

The invention belongs to the technical field of biology, and particularly relates to a porcine epidemic diarrhea virus broad-spectrum neutralizing antibody and application thereof. A porcine epidemic diarrhea virus broad-spectrum neutralizing antibody 6G2 is screened, the virus neutralizing activity of the antibody is evaluated through a cell model, and it is found that the antibody can effectively block infection of G1a, G1b, G2a, G2b, G2c and S-INDEL (porcine epidemic diarrhea virus) gene subtype strains with high epidemic rate on cells. Further, animal infection experiments of pigs prove that the neutralizing antibody provided by the invention has good prevention and treatment effects on the porcine epidemic diarrhea virus, and has important significance on improving the capability of preventing and controlling porcine epidemic diarrhea in a pig farm.

The invention discloses an indirect ELISA (enzyme-linked immuno sorbent assay) detection method of a porcine epidemic diarrhea virus antibody SIgA, and belongs to the technical field of animal antibody detection. In order to solve the problems of limited detection range and single immune evaluation dimension of a porcine epidemic diarrhea virus detection method in the prior art, the invention provides an indirect ELISA detection method of a porcine epidemic diarrhea virus antibody SIgA, and the detection method adopts a pichia pastoris eukaryotic secretion platform to prepare a high-activity PEDV S1 antigen; meanwhile, a secreting type IgA antibody is used as a detection target; the method has the advantages of high detection sensitivity, low background noise, better variant compatibility and practical popularization value, and provides a clinical and industrial-level solution for maternal immunity monitoring, piglet passive protection evaluation and vaccine immunity strategy optimization.

The invention relates to the technical field of veterinary biological products, and particularly provides a porcine epidemic diarrhea virus S protein and related products. The amino acid sequence of the porcine epidemic diarrhea virus S protein fragment is as shown in SEQ ID NO.4. The trimer S protein is constructed by fusing a trimer tag at the C terminal of a protective antigen in a truncated form of the porcine epidemic diarrhea virus S protein. After expression and purification, immunogenicity is judged through animal evaluation, and it is verified that the vaccine prepared from the trimer S protein has the advantages of being high in safety, good in immunogenicity, stable in batches, capable of inducing piglets to generate good immunoreaction and the like, and porcine epidemic diarrhea can be effectively treated or prevented.

The invention discloses a porcine epidemic diarrhea virus S protein monoclonal antibody and application thereof. Specifically, two monoclonal antibodies GC42G2-1 and GC33D8-1 which have broad-spectrum binding characteristics with the porcine epidemic diarrhea virus S protein are obtained firstly, the GC42G2-1 antibody has good universality and can have strong affinity with different strains, and the GC33D8-1 antibody can be used for binding the porcine epidemic diarrhea virus S protein in a broad-spectrum manner. The GC42G2-1 can be independently used, and also can be matched with a polyclonal antibody or GC33D8-1 to be used for sandwich ELISA (enzyme-linked immunosorbent assay) so as to detect and quantify PEDV (porcine epidemic diarrhea virus). Secondly, the two monoclonal antibodies are respectively used as a coating antibody and a detection antibody, a double-antibody sandwich ELISA method is established, and the double-antibody sandwich ELISA method is applied to PEDV vaccine S protein quantification, has better parallelism compared with other antibodies, and can be widely applied to detection of different strains. The method disclosed by the invention has the advantages of high sensitivity, high accuracy, high repeatability and high flux, and can be used for the development of blocking ELISA (Enzyme-Linked Immunosorbent Assay), capturing ELISA, indirect ELISA, competitive ELISA and detection of porcine epidemic diarrhea viruses by colloidal gold test strips.

The invention provides a porcine epidemic diarrhea virus G2c subtype strain and application thereof. According to the invention, the porcine epidemic diarrhea virus G2c subtype HN2501 strain is separated from porcine intestinal tissues and is subjected to passage purification, and the microbial preservation number of the porcine epidemic diarrhea virus G2c subtype HN2501 strain is CGMCC (China General Microbiological Culture Collection Center) No.46417. The isolated strain can stably proliferate on passage cells to generate typical cytopathy. The G2c subtype isolated strain of the porcine epidemic diarrhea virus has excellent immunogenicity; the vaccine prepared from the isolated strain can induce piglets to generate high-level neutralizing antibodies; meanwhile, the ELISA kit prepared from the isolated strain can be used for well detecting the porcine epidemic diarrhea virus antibody in serum.

The invention provides a subunit vaccine composition for porcine epidemic diarrhea, porcine delta coronal and porcine rotavirus as well as a preparation method and application of the subunit vaccine composition, and is characterized in that 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 porcine epidemic diarrhea virus S protein, porcine delta coronavirus S protein and porcine rotavirus VP8 protein. 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 application belongs to the technical field of biotechnology, and particularly relates to a porcine epidemic diarrhea virus recombinant strain, a construction method and application. The S gene sequence of the existing epidemic porcine epidemic diarrhea virus PEDV CH / SX / 2016 strain is replaced by the S2015 gene sequence shown in SEQ ID NO. 1 to construct a PEDV CH / SX / 2016 strain infectious clone. A porcine epidemic diarrhea virus recombinant strain rCH / SX / 2016-S 2015 is successfully rescued by transfecting cells with a plasmid containing the infectious cDNA shown in SEQ ID NO. 4. The problem that the original strain cannot be successfully rescued by reverse genetics engineering is solved, and the recombinant virus strain rCH / SX / 2016-S 2015 has an activity comparable to that of the original strain, and can be used for preparing a PEDV diagnostic reagent and / or vaccine.

The invention provides an application of brucein A in preparation of an anti-porcine epidemic diarrhea virus drug, and belongs to the technical field of chemical medicines. Experimental results show that the brucein A has remarkable inhibiting and blocking effects on a-1 ribosomal Frameshift process of a porcine epidemic diarrhea virus (PEDV), can effectively inhibit replication and proliferation of the PEDV, and has an inhibiting effect on a plurality of common strains of the PEDV; the brucein A can be used for preparing a medicine for resisting PEDV (porcine epidemic diarrhea virus) and a medicine for preventing and treating porcine epidemic diarrhea caused by PEDV, and the medicine is a wide-spectrum antiviral medicine, is insensitive to variation of PEDV and has a wide application prospect.

The invention discloses a primer probe combination for simultaneously detecting African swine fever virus, porcine epidemic diarrhea virus and porcine reproductive and respiratory syndrome virus, a kit and application thereof. The primer probe combination consists of a first primer probe group for detecting African swine fever virus, a second primer probe group for detecting porcine epidemic diarrhea virus and a third primer probe group for detecting porcine reproductive and respiratory syndrome virus. The primer probe combination for simultaneously detecting the African swine fever virus, the porcine epidemic diarrhea virus and the porcine reproductive and respiratory syndrome virus, provided by the invention, is high in sensitivity and good in specificity, and can be effectively used for detecting the African swine fever virus, the porcine epidemic diarrhea virus and the porcine reproductive and respiratory syndrome virus.

The application discloses a RT-MIRA-LFD kit for detecting PEDV and belongs to the technical field of molecular biology. The application firstly provides a RT-MIRA-LFD primer probe combination for rapidly detecting porcine epidemic diarrhea virus, which comprises an upstream primer shown as SEQ ID NO. 5, a downstream primer shown as SEQ ID NO. 7 and a probe shown as SEQ ID NO. 9; secondly, the application provides a kit and a detection method for rapidly detecting porcine epidemic diarrhea virus. The application designs the primer and the probe for detecting porcine epidemic diarrhea virus based on the RT-MIRA-LFD technology, and designs the RT-MIRA-LFD detection method. The detection method has the advantages of high sensitivity and high specificity, short detection time and visual results, and provides a new experience method for rapid detection of porcine epidemic diarrhea virus.

This invention belongs to the field of biotechnology, specifically relating to a broad-spectrum neutralizing antibody against porcine epidemic diarrhea virus (PEDV) and its applications. This invention screened a broad-spectrum neutralizing antibody against PEDV, 6G2. Cell model evaluation revealed that the antibody effectively blocked infection of cells by several highly prevalent PEDV genotypes: G1a, G1b, G2a, G2b, G2c, and S-INDEL. Further animal infection experiments in pigs demonstrated that the neutralizing antibody provided by this invention has good preventive and therapeutic effects against PEDV, which is of great significance for improving the ability of pig farms to control PEDV.

The invention discloses a porcine epidemic diarrhea and porcine rotavirus (G9 type) bigeminy attenuated vaccine and a preparation method thereof, and relates to the technical field of veterinary biological products. According to the present invention, dominant genotype strains such as PEDV G2 type KQ01 strain (the preservation number is CCTCC NO: V202005) and PoRV G9 type CH09 strain (the preservation number is CCTCC NO: V202191) are adopted as parent strains, a cell continuous passage method is adopted to carry out passage attenuation, and the virus content, the purity, the specificity, the hereditary stability, the safety and the immunogenicity are systematically inspected during the passage process; and then, carrying out proliferation process research on passage attenuated strains PEDV HB17-R and PoRV JS01-R in a bioreactor, harvesting a cell culture, mixing the two antigens according to a proper ratio, adding a proper amount of a stabilizer, and carrying out a freeze vacuum drying process to obtain the high-purity PoRV-HB17-R / PoRV-HB17-R / PoRV-HB17-R / PoRV-HB17-R / PoRV-JS01-R composite antigen. The bigeminy attenuated vaccine is highly matched with current dominant prevalent strains (PEDV G2 type and PoRV G9 type), is based on a suspension culture technology and has both safety and high efficiency.

Use of MMPD for the preparation of a medicament for inhibiting porcine epidemic diarrhea virus (PEDV) infection in vitro.

The invention relates to the technical field related to immunological detection, in particular to a mouse anti-porcine epidemic diarrhea virus monoclonal antibody, an immunogen and application thereof. The amino acid sequences of complementary determining regions CDR1, CDR2 and CDR3 of a heavy chain variable region of the monoclonal antibody or the antigen binding fragment of the monoclonal antibody are respectively as follows: GFSLSTSGVV, RIDPSDSYT and ARDREKSDNSGYFDY, and the amino acid sequences of complementary determining regions CDR1, CDR2 and CDR3 of a light chain variable region of the monoclonal antibody or the antigen binding fragment of the monoclonal antibody are respectively as follows: RASKSISKYLA, EASSLES and LQHTPKKNK. The monoclonal antibody can be specifically combined with nucleocapsid proteins (N proteins) of all subtypes of the porcine epidemic diarrhea virus, and a porcine epidemic diarrhea virus detection kit prepared from the monoclonal antibody has the advantages of high sensitivity, strong specificity, wide detection range, short detection time and the like.

A composition comprising Bidens pilosa or an active compound isolated from the Bidens pilosa for use in treating, inhibiting and / or decreasing the presence of pathogenic gut microbiota in an animal in need thereof is disclosed. The pathogenic gut microbiota are at least one selected from the group consisting of Brachyspira, porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis coronavirus (TGEV) and Rotavirus (RV). The composition is also for use in treating, inhibiting and / or decreasing occurrence of swine diarrhea, swine dysentery, swine viral infection and increasing the percentage of swine carcass and bone weight. In one embodiment, the composition is for use in treating, inhibiting and / or decreasing occurrence of swine dysentery associated with Brachyspira.

The present invention provides a porcine epidemic diarrhea virus S protein mutant, a nucleic acid molecule, a subunit vaccine and applications thereof; compared with the wild-type porcine epidemic diarrhea virus S protein, the porcine epidemic diarrhea virus S protein mutant has the 1076th amino acid, the 1077th amino acid, the 1016th amino acid and the 1017th amino acid all mutated into proline. The present invention constructs a stable natural trimer that maintains the prefusion conformation of the S protein, obtains a safe and highly effective PEDV subunit vaccine, and can improve the use effect and immunogenicity of the porcine epidemic diarrhea vaccine; moreover, compared with the wild-type S protein, the porcine epidemic diarrhea virus S protein mutant in the present invention has a higher expression level.

The invention relates to the technical field of mRNA vaccine preparation, and particularly discloses a porcine epidemic diarrhea virus mRNA vaccine and a preparation method thereof. The mRNA vaccine is prepared by adopting cow milk source exosome as a carrier and wrapping mRNA encoding PEDV-S protein in the carrier through an ultrasonic loading method. According to the preparation method, the encapsulation efficiency of mRNA and the stability of the vaccine are remarkably improved, the prepared vaccine can more efficiently promote antigen expression, stronger cellular immunity and humoral immunity are induced in a mouse body, and a safe and efficient novel vaccine candidate is provided for preventing and controlling porcine epidemic diarrhea.

The present application relates to the field of biological medicine, in particular to a kind of muscone in the application of viral inhibitor.The viral inhibitor of the present application is used to inhibit the replication of Japanese encephalitis virus and / or porcine epidemic diarrhea virus.The present application finds that muscone has no cytotoxicity, and does not directly inactivate virus, but produces antiviral effect on Japanese encephalitis virus by inhibiting NLRP3 / Caspase-1, Caspase-9 / Caspase-3 / Bax / BCL-2, JNK / ERK / P38 and other signal pathways, and inhibits porcine epidemic diarrhea virus invasion into cells by reducing caveolin level.The antiviral effect is remarkable, the mechanism of action is clear, the selectivity is high, and the safety is better.

The invention discloses biomedical application of 3, 29-dibenzoyl karounitriol, and particularly relates to application of 3, 29-dibenzoyl karounitriol in inhibition of physiological activities of coronavirus and application of 3, 29-dibenzoyl karounitriol in preparation of drugs for preventing and treating coronavirus infectious diseases. The invention proposes that the triterpenoid 3, 29-dibenzoyl karounitriol can inhibit the replication of porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine delta coronavirus for the first time; experimental data show that the compound can effectively prevent and treat porcine epidemic diarrhea and has a good application prospect in the domestic pig breeding industry.

This invention provides a method and system for preventing the spread of porcine epidemic diarrhea virus (PEDV), comprising: periodically collecting the first total feed intake of a target pig pen, and generating a feed intake time-series curve of the target pig pen based on the first total feed intake; if the feed intake time-series curve identifies a continuous decrease in total feed intake within a preset time period, generating a meal control instruction for the target pig pen based on preset rules; and sending the meal control instruction to the feeding controller corresponding to the target pig pen, so that the feeding controller delivers feed and water according to the meal control instruction. This invention improves upon the shortcomings of existing technologies in terms of timeliness and accuracy.

The invention relates to a fluorescent RT-ERA detection primer probe composition for porcine epidemic diarrhea virus and application, and belongs to the technical field of virus detection. The primer probe composition comprises an upstream primer, a downstream primer and a probe, and the sequence of the upstream primer is shown as SEQ ID NO: 15; the sequence of the downstream primer is as shown in SEQ ID NO: 16; the sequence of the probe is as shown in SEQ ID NO: 3. The invention also provides an application of the fluorescent RT-ERA detection primer probe composition for the porcine epidemic diarrhea virus in preparation of a product for detecting the porcine epidemic diarrhea virus. The primer probe composition can realize rapid diagnosis of the porcine epidemic diarrhea virus, is high in sensitivity, simple and convenient to operate and good in stability, and provides powerful technical support for basic prevention and control of the porcine epidemic diarrhea virus and other important epidemic diseases.

The application discloses a porcine epidemic diarrhea disease vaccine composition and a preparation method and application of the vaccine composition. The inactivated vaccine can effectively protect against the attack of epidemic virulent strains, has good cross-protection, and can effectively protect a pig group against porcine epidemic diarrhea and improve the productivity of the pig group.

This invention relates to the field of biology, specifically providing a hybridoma cell line targeting the S1 gene of the G2c variant of porcine epidemic diarrhea virus (PEDV) and the specific antigenic epitope recognized by the antibody secreted by this cell line. 680 SGQLLAFKNVTSGAVYSV 697 This invention also discloses the heavy and light chain variable region sequences of the antibody. The hybridoma cell line exhibits high specificity and stability, and the antibody it secretes can specifically recognize the S1 protein of the PEDV G2c variant. This provides key biomaterials and technical support for the rapid diagnosis, vaccine development, and pathogenic mechanism research of the PEDV G2c variant, and has significant application value in the field of livestock disease prevention and control.

The invention belongs to the technical field of biological detection, and particularly relates to a porcine epidemic diarrhea recombinant rS1 antigen as well as a preparation method and application thereof. A coding sequence of the recombinant rS1 antigen sequentially comprises the following elements from an N terminal to a C terminal: a signal peptide, PEDV S1 protein and a trimer tag, and the elements are connected with one another through linkers. According to the recombinant rS1 antigen, a signal peptide capable of assisting S1 protein secretion is added to the upstream of an S gene to assist soluble expression of S1 protein in periplasmic space, so that the protein solubility is enhanced; a trimer tag GCN4 is added to the downstream to assist correct folding of S1 in the expression process, so that the protein solubility is further enhanced, and correct expression of conformation is realized.

The invention relates to the technical field of biology, and discloses a construction method of a cell line for expressing porcine epidemic diarrhea virus S1 protein.The method comprises the steps that a nucleotide sequence of the PEDV S1 protein is inserted into a plasmid, and the plasmid carrying nucleotide of the PEDV S1 protein is obtained; then transfecting the plasmid carrying the nucleotide of the PEDV S1 protein into CHO-K1, and culturing to obtain a cell line for expressing the PEDV S1 protein; the nucleotide sequence of the PEDV S1 protein is shown as SEQ ID NO: 1, the cell line capable of stably expressing the PEDV S1 protein is obtained through the design, and in addition, the invention further discloses the cell line capable of expressing the PEDV S1 protein, application of the cell line and an ELISA detection method.

The invention belongs to the technical field of veterinary biological products, and relates to an mRNA freeze-dried vaccine of a porcine epidemic diarrhea virus as well as a preparation method and application of the mRNA freeze-dried vaccine. When the mRNA vaccine is prepared, firstly, the S gene sequences of the porcine epidemic diarrhea virus GIIa type and the porcine epidemic diarrhea virus GIIc type which are separated and stored in a laboratory are selected; the preparation method comprises the following steps: firstly, carrying out codon optimization, then respectively connecting with 5 'UTR, 3' UTR, a termination codon and a polyA tail nucleic acid sequence in series, connecting to a vector, carrying out in-vitro template transcription, carrying out in-vitro transcription, capping and purifying to obtain mRNA, and finally, encapsulating and freeze-drying to prepare the mRNA vaccine. The invention further provides application of the vaccine to immunization of pigs, cellular immunity and humoral immunity in the pigs can be stimulated, the immunogenicity and protective efficacy of the vaccine are superior to those of commercially available bivalent live vaccines, and the vaccine can be used for preventing porcine diarrhea caused by infection of currently epidemic PEDV GIIa and GIIc type strains.

The present invention discloses a porcine epidemic diarrhea virus strain, a nanobody and an antibody cocktail. The porcine epidemic diarrhea virus strain is the porcine epidemic diarrhea virus PEDV-JSXH-2021 strain, whose deposit number is CGMCC No.46088, and was deposited in the General Microbiology Center of the China Culture Collection Administration on August 14, 2024. The nanobody is obtained by screening the porcine epidemic diarrhea virus PEDV-JSXH-2021 strain using yeast two-hybrid technology. The present invention obtains specific nanoantibodies targeting PEDV through yeast two-hybrid technology. The obtained nanoantibodies have neutralizing activity against PEDV and can effectively exert anti-PEDV effects. These nanoantibodies are non-toxic in vitro and can neutralize PEDV infection by binding to different epitopes of the spike protein, providing a theoretical basis and new research direction for the development of VHH-based drugs to treat and prevent PEDV-induced diarrhea.

The invention relates to the technical field of molecular biology and immunology, in particular to a porcine epidemic diarrhea virus-like particle subunit vaccine and a preparation method thereof. The invention provides a porcine epidemic diarrhea virus-like particle subunit vaccine. The vaccine is formed by assembling partial fragments of porcine epidemic diarrhea PEDV S protein and virus-like particles. Compared with a traditional subunit vaccine, the porcine epidemic diarrhea virus-like particle vaccine has the advantages that the immune efficacy is greatly improved, compared with an inactivated vaccine, the porcine epidemic diarrhea virus-like particle vaccine has a better immune effect, has no pathogenicity to animals such as mice and the like, and can induce high-level neutralizing antibodies in animal bodies.