18 results about "Acute diarrhea" patented technology

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 discloses an antibody for detecting porcine acute diarrhea syndrome coronavirus, a fluorescence immunochromatography test strip and application of the antibody and the fluorescence immunochromatography test strip, and aims to overcome the defects that an existing SADS-CoV detection technology is low in sensitivity, complex to operate and incapable of being applied on site. The method comprises the following steps: screening high-specificity anti-S1 protein monoclonal antibodies (such as 1B8 and 3C9); a quantum dot labeled antibody is combined with a chromatographic structure containing a T line (S1 protein) and a C line (SPA protein); the kit provided by the invention has the advantages that by optimizing the formula of the buffer solution, the technical effects of high-sensitivity detection (100 TCID50 / mL), on-site rapid visual interpretation and zero cross reaction are realized, the technical bottleneck of on-site rapid and accurate diagnosis is overcome, the operation is simple and convenient, and the kit is suitable for on-site screening of clinical samples.

The invention discloses a multiplex nucleic acid detection kit for swine acute diarrhea pathogens and application of the multiplex nucleic acid detection kit. The kit comprises an RAA (Reverse Amplification) primer pair, a specific crRNA (Ribonucleic Acid), a Cas12a protein, an NEBbuffer r2.1 and an ssDNA (Deoxyribose Nucleic Acid) reporter molecule, wherein the RAA primer pair, the specific crRNA, the Cas12a protein, the NEBbuffer r2.1 and the ssDNA reporter molecule are used for detecting the porcine epidemic diarrhea virus (PEDV), the porcine rotavirus (PoRV) and the porcine sapelovirus (PSV). According to the method, reverse transcription, amplification and Cas12a detection processes are coupled through an integrated system or a continuous reaction mode, the whole process is completed under the condition of constant temperature (37 DEG C), thermal cycle equipment is not needed, and only a constant-temperature heating device is needed. After the reaction is finished, a result can be observed through a portable blue light detector or naked eyes, so that on-site real-time visual interpretation is realized.

The invention belongs to the technical field of biological medicines, and provides an S protein ancestor sequence AS mRNA of a porcine acute diarrhea syndrome coronavirus, the ancestor sequence of an SADS-CoV Spike protein is constructed through phylogenetic analysis, and the sequence aims at the existing virus and also aims at the evolution pedigree of the virus; the sequence design is used for converting target defense aiming at a single pathogen into active intervention on a virus evolution trajectory; and the prepared AS mRNA vaccine has important significance on feeding safety of animals and prevention and control of zoonosis diseases.

The present application belongs to the technical field of biological industry, and particularly relates to a spike protein antigen epitope peptide of porcine acute diarrhea syndrome coronavirus, a monoclonal antibody thereof and application. The present application provides an antibody or an antigen binding fragment thereof, a nucleotide sequence of a heavy chain variable region of the antibody or the antigen binding fragment thereof is shown as SEQ ID NO. 2; a nucleotide sequence of a light chain variable region of the monoclonal antibody is shown as SEQ ID NO. 4. A minimum binding epitope amino acid sequence is shown as SEQ ID NO. 22, and the antigen epitope is highly conserved in various different subtypes of SADS-CoV. The antibody can be applied to basic research on a SADS-CoV invasion molecular mechanism, and provides good scientific research materials for application research of SADS-CoV detection and treatment.

The present invention belongs to the field of biotechnology and specifically relates to a latex dual immunochromatographic test strip for rapid identification and detection of porcine acute diarrhea syndrome coronavirus (SADS-CoV) and porcine transmissible gastroenteritis coronavirus (TGEV), and its preparation method. The test strip comprises a PVC base, a sample pad, a conjugation pad, a nitrocellulose membrane, and an absorbent pad. The nitrocellulose membrane is provided with a quality control line C, test lines T1, and test lines T2. The conjugation pad is sprayed with latex microspheres labeled with monoclonal antibodies 1B7 against the TGEV N protein and 6E8 against the SADS-CoV N protein. The test line T1 is coated with the anti-SADS-CoV N protein monoclonal antibody 6E8, the test line T2 is coated with the anti-TGEV N protein monoclonal antibody 2A9, and the quality control line C is coated with goat anti-mouse IgG. This test strip has the advantage of being a dual-link test strip, specifically enabling differential diagnosis of TGEV and SADS-CoV, whose clinical symptoms are easily confused.

The invention discloses a porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody and application thereof, and belongs to the technical field of biology. According to the invention, capsid protein of porcine acute diarrhea syndrome coronavirus SADS-CoV is taken as a detection marker, recombinant capsid protein of SADS-CoV is expressed through an eukaryotic expression system, a monoclonal antibody of anti-capsid protein is prepared by using a hybridoma technology, and then the SADS-CoV colloidal gold rapid diagnosis test strip is prepared by using the monoclonal antibody and a colloidal gold test strip evaluation system. The porcine acute diarrhea syndrome coronavirus capsid protein monoclonal antibody provided by the invention is used for preparing an immunochromatography detection test strip, can be used for rapidly detecting porcine acute diarrhea syndrome coronavirus, and is simple, convenient, easy in result judgment and suitable for field.

The composition may be used therapeutically or prophylactically and is directed toward a cluster of diarrhea-causing pathogens which cause illness or death in animals, including dogs and cats. It is prepared from a powdered egg preparation and powdered protein matrix, such as bovine colostrum. The eggs are collected from hens which have been immunized with the relevant pathogens or toxins. When the matrix includes colostrum, the powdered colostrum is derived from non-hyperimmune cattle. The vaccination strategy includes the use of antibody cross-reactivity between toxins or pathogens which cause diarrhea. For some diseases, including canine parvo, the clinical improvement using this therapeutic exceeds the standard of care. Instead of a pharmaceutical product, this composition is an orally administered food product with the same safety profile as eggs and milk.

The application belongs to the technical field of veterinary medicine, and particularly relates to an adenovirus vector recombinant pig acute diarrhea syndrome coronavirus vaccine. The application constructs a replication-defective adenovirus vector vaccine (rAd-SADS-S) expressing a full-length SADS-CoV S protein, realizes immune induction of high-efficiency humoral and cellular immunity, and provides passive protection for milk mice through maternal antibody transmission, and overcomes the defects of no available vaccine for SADS-CoV, low protection rate in existing research, and biological safety risk.

The invention belongs to the technical field of veterinary medicine, and particularly relates to an adenovirus vector recombinant porcine acute diarrhea syndrome coronavirus vaccine. According to the invention, by constructing a replication-deficient adenovirus vector vaccine (rAd-SADS-S) expressing the full length of SADS-CoV S protein, efficient body fluid and cell immunity is induced by immunity, passive protection is provided for suckling mice through maternal antibody transfer, and the defects that no SADS-CoV vaccine is available and the existing research protection rate is low and the biological safety risk is caused are overcome.

The invention discloses a primer group for simultaneously detecting porcine acute diarrhea syndrome coronavirus and bat HKU2 coronavirus, and belongs to the technical field of biology. The primer combination disclosed by the invention can be used for simultaneously detecting the porcine acute diarrhea syndrome coronavirus (SADS-CoV) and the bat HKU2 coronavirus, and the lowest limit of detection (LOD) is 20 copies per reaction; the primer provided by the invention is combined with three gene conserved regions of SADS-CoV and HKU2 coronavirus, and has relatively high specificity. The method has high reference value for nucleic acid diagnosis of SADS-CoV and HKU2 coronavirus, and is suitable for popularization and application.

The invention belongs to the technical field of traditional Chinese medicine. The invention relates to a traditional Chinese medicine composition, in particular to a traditional Chinese medicine composition for clearing heat, resolving dampness, relieving spasm and stopping diarrhea and a preparation method and application thereof. The invention provides a traditional Chinese medicine composition, which is prepared from the following raw materials in parts by weight: 4 to 12 parts of herba andrographitis, 2 to 8 parts of rhizoma coptidis, 2 to 8 parts of quassia wood, 1 to 6 parts of radix aucklandiae, 2 to 8 parts of radix pulsatillae, 2 to 8 parts of radix paeoniae alba, 2 to 8 parts of semen plantaginis, 2 to 8 parts of red-knees herb and 1 to 6 parts of liquorice root. Researches show that the traditional Chinese medicine composition has a remarkable treatment effect on model mice with acute enteritis induced by escherichia coli, can play a remarkable curative effect after being administrated once, takes effect quickly, and meets the clinical treatment requirements of relieving urgency and stopping diarrhea. According to the traditional Chinese medicine composition, the specific types and proportions of the medicines are controlled, the synergistic effect of the components is improved, pathogenic microorganism infection can be inhibited, the intestinal function can be adjusted, the effect of treating both symptoms and root causes is achieved, and the traditional Chinese medicine composition has remarkable social benefits and clinical application value.

The invention belongs to the technical field of veterinary medicine, and relates to a recombinant pseudorabies virus strain for expressing porcine acute diarrhea syndrome coronavirus S protein as well as a construction method and application of the recombinant pseudorabies virus strain. The recombinant pseudorabies virus strain is obtained by rescuing recombinant pseudorabies virus infectious clone for expressing S protein of porcine acute diarrhea syndrome coronavirus; the recombinant pseudorabies virus infectious clone comprises five Fosmid cosmiles, the five Fosmid cosmiles contain gene segments, namely Fos-1, Fos-2-dTK, Fos-3-dTK, Fos-4 and Fos-5-SADS-S, which are used for rescuing the pseudorabies virus, are mutually overlapped and cover a pseudorabies virus genome, and the Fos-5-SADS-S contains a porcine acute diarrhea syndrome coronavirus S protein gene expression cassette. After a mouse is inoculated with the recombinant pseudorabies virus strain, remarkable PRV and SADS-CoV specific antibodies and neutralizing antibodies can be induced to be generated. An immunized mouse can resist the lethal dose attack of a PRV virulent strain, and a suckling mouse produced by the immunized mouse can resist the lethal dose attack of SADS-CoV.

The invention belongs to the technical field of biological medicines, and particularly relates to a composition for improving intestinal dysfunction and a preparation method and application thereof. The composition is prepared from the following raw materials in parts by weight: 18 to 25 parts of collagen peptide, 14 to 20 parts of yeast polypeptide, 10 to 16 parts of betaine, 7 to 15 parts of emblic leafflower fruit extract, 6 to 12 parts of olive extract, 9 to 14 parts of xylooligosaccharide and 3 to 5 parts of yeast beta-glucan. The amino acid sequence of the collagen peptide is as shown in SEQ ID NO. 1. By taking the composition prepared by the invention, acute diarrhea and constipation can be quickly relieved, intestinal inflammation can be improved, and the composition can enduringly act on the internal environment of intestinal tracts, so that deep and enduringly improvement of intestinal dysfunction is realized, and the composition has a wide application prospect.

The invention discloses separation, identification and application of a porcine bocavirus cell adapted strain, and belongs to the technical field of gene engineering. The preservation number is CGMCC (China General Microbiological Culture Collection Center) No.46413, and the strain can be stably proliferated in passage cells of porcine kidney cells (PK) and can induce a remarkable cytopathic effect. Virus separation and identification are completed through metagenome sequencing, transmission electron microscope observation and indirect immunofluorescence technologies, and a phylogenetic tree is constructed based on a VP1 gene sequence. The invention also establishes a piglet infection model, and proves that the porcine bocavirus cell adapted strain CNH-2024 can cause acute diarrhea and lung pathological injury. The invention also provides a specific fluorescent quantitative PCR primer combination, a vaccine preparation method and a diagnostic kit. The porcine bocavirus cell adapted strain CNH-2024 obtained by separation and identification provided by the invention provides an important technical basis for porcine bocavirus pathogenic mechanism research, vaccine development and clinical diagnosis.

The composition may be used therapeutically or prophylactically and is directed toward a cluster of diarrhea-causing pathogens which cause illness or death in animals, including dogs and cats. It is prepared from a powdered egg preparation and powdered protein matrix, such as bovine colostrum. The eggs are collected from hens which have been immunized with the relevant pathogens or toxins. When the matrix includes colostrum, the powdered colostrum is derived from non-hyperimmune cattle. The vaccination strategy includes the use of antibody cross-reactivity between toxins or pathogens which cause diarrhea. For some diseases, including canine parvo, the clinical improvement using this therapeutic exceeds the standard of care. Instead of a pharmaceutical product, this composition is an orally administered food product with the same safety profile as eggs and milk.

The invention relates to a monoclonal antibody of porcine acute diarrhea syndrome coronavirus S1-CTD protein and application of the monoclonal antibody. According to the invention, S1-CTD protein is expressed in BL21 (DE3) cells by using a prokaryotic expression system, the purified S1-CTD protein is used as an immunogen, a BALB / c mouse is immunized by an immunological method, and the monoclonal antibody for resisting SADS-CoV S1-CTD protein is prepared through cell fusion. Two rounds of truncation are carried out by using an overlapping polypeptide method, the antigen epitope is identified by indirect ELISA (Enzyme-Linked Immunosorbent Assay) and Dot-ELISA, and the result shows that the linear B cell epitope region of the S1-CTD protein is PGFVLRVGRG. The monoclonal antibody and the B cell epitope lay a foundation for preparing a reagent for diagnosing or preventing SADS-COV.

This invention utilizes an insect-baculovirus expression system to prepare the recombinant protein SADS-CoV-S of porcine acute diarrhea syndrome coronavirus, which is then used to prepare egg yolk antibodies. By optimizing the composition of the subunit vaccine composition for immunizing laying hens, IL-2 and ZnCl2 solution are added as immunostimulants to the conventional adjuvant, increasing the antibody concentration in the egg yolk antibody extract and improving the neutralizing antibody titer and protective efficacy. When IL-2 and zinc chloride aqueous solution are added to the immunization composition simultaneously, the resulting egg yolk antibody aqueous solution exhibits the highest protein concentration, and at the same protein concentration, the egg yolk antibody has the highest neutralizing titer. Compared to the immunization composition using Freund's complete adjuvant alone, the neutralizing titer is increased by more than 60 times.