13 results about "Diarrheal diseases" patented technology

The invention belongs to the technical field of virus detection, and particularly relates to a porcine delta coronavirus N protein monoclonal antibody and a latex immunochromatography test strip containing the antibody. According to the invention, a prokaryotically expressed PDCoV N protein is used as an immunogen to immunize a mouse, and a specific monoclonal antibody 2G3 and a specific monoclonal antibody 9C4 are screened through cell fusion and a subcloning technology. The test strip comprises a PVC (polyvinyl chloride) bottom plate, a sample pad, a combination pad, a nitrocellulose membrane (NC membrane) and a water absorption pad, a quality control line C and a detection line T are respectively arranged on the NC membrane. The test strip can specifically recognize PDCoV, the lowest detection limit can reach 103.0 TCID50 / mL, the test strip has no cross reaction with other clinical common diarrhea viruses, the result interpretation only needs 15 min, and the test strip has the advantages of being rapid, sensitive, easy and convenient to operate and the like.

The present invention relates to the technical field of virus detection, and discloses a sewage diarrhea virus typing detection system, which comprises a pathogen enrichment unit, an improved polyethylene glycol precipitation method is adopted to carry out pathogen enrichment on a sewage sample; and the nucleic acid extraction unit is used for extracting nucleic acid in the enriched sample by using a magnetic bead extraction method. According to the typing detection system for the diarrhea viruses in the sewage, simultaneous detection and typing of various diarrhea viruses such as norovirus, rotavirus, adenovirus, astrovirus, parareovirus and fiveleaf virus in the sewage can be realized, the detection range is widened, powerful support is provided for comprehensive monitoring of virus pollution conditions in the sewage, and the system is suitable for popularization and application. According to the present invention, the multiple fluorescent RT-PCR technology is adopted, and the efficient sample enrichment and nucleic acid extraction method is combined, such that the detection sensitivity is improved, the low-concentration viral nucleic acid can be detected, and the potential viral threat is not missed;

The invention discloses an epegRNA (Ribonucleic Acid) primer pair for detecting an Indel marker related to porcine diarrhea virus resistance and application of the epegRNA primer pair. According to the epegRNA sequence provided by the invention, accurate AACA insertion mutation can be realized on a sequence of which the pig genome position is ChrX: 56448702 through pilot editing; the A / AAACA mutation can be used as a molecular marker to be applied to breeding for resisting porcine epidemic diarrhea.

The application discloses an ELISA method for detecting porcine epidemic diarrhea virus antibody SIgA and belongs to the technical field of biology. The method comprises the steps of taking S1 protein as a coating antigen and detecting porcine epidemic diarrhea virus antibody SIgA in a sample to be detected, wherein the amino acid sequence of the S1 protein is a truncated sequence from the 19th to the 719th. The application preliminarily establishes an indirect ELISA method for detecting porcine epidemic diarrhea virus specific SIgA antibodies in serum, intestinal tissue and milk by optimizing conditions, and determines that the optimal antigen coating concentration and coating time are 0.25 mu g / ml and 4 DEG C overnight 12h-16h respectively, and the optimal detection conditions are determined. The method for detecting PEDV specific SIgA in serum, intestinal tissue and milk established in the research is simple and rapid, has good sensitivity and specificity, and provides a basis and a detection standard for evaluating the mucosal immune level of PEDV.

The invention provides a composite inactivator for inactivating bovine diarrhea virus, an inactivation method of bovine diarrhea virus and application thereof, and relates to the technical field of virus inactivation, the composite inactivator comprises a DMEM solution containing methylene blue and inorganic salt; wherein the inorganic salt comprises sodium chloride, magnesium chloride and potassium chloride; the inactivation method of the bovine diarrhea virus comprises the following steps: under light treatment, inactivating the bovine diarrhea virus through a composite inactivator containing methylene blue to obtain an inactivated bovine diarrhea virus liquid. The technical problems that in the prior art, inactivation of the bovine diarrhea virus has biological harm, consumed time is long, and operation is complex are solved, the technical effects of being simple, convenient, feasible, safe, stable, free of biological harm and short in consumed time are achieved, meanwhile, the pollution resistance of the product is effectively improved, and the shelf life of the product is prolonged.

The application discloses a kind of pig multiple diarrhea virus detection method and application based on tNGS technology.The method realizes the accurate detection of pig epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine delta coronavirus by tNGS technology.The primer set involved is used to amplify the S1 and M genes of PEDV, the N gene of TGEV and the M gene of PDCoV.The application also provides a kit comprising the primer set and a corresponding tNGS detection method.Clinical sample verification shows that the method is completely consistent with the qPCR detection results, has high sensitivity, strong specificity, can simultaneously identify mixed infection and other advantages, and has high detection throughput, controllable cost, is suitable for accurate diagnosis, epidemiological monitoring and prevention and control of diarrhea virus in pig farm, and has wide application prospect.

The invention discloses and relates to an antiviral RNA molecule capable of inhibiting porcine diarrhea virus and a design, preparation and toxicity detection method, the antiviral RNA molecule is PEDV-mhRNA, the antiviral RNA molecule is formed by connecting a plurality of siRNAs of S protein coding genes of targeted PEDV in series, an RNAi medicine capable of being used for preventing and treating PEDV is provided for domestic pig breeding, and the antiviral RNA molecule can be used for preventing and treating the PEDV. According to the present invention, the replication and the proliferation of the PEDV in the vero cell can be significantly inhibited, the method can be used for the prevention and the treatment of the PEDV, the RNA is adopted as the anti-virus factor, the characteristics of strong specificity, good anti-virus effect, no harm to the virus host and other organisms and the like are provided, the applicability is strong, the economic benefits of the breeding industry can be improved, and the promotion and application significance is provided.

The invention discloses a primer probe composition, a kit and a method for detecting multiple cattle and sheep diarrhea pathogens based on nucleic acid mass spectrometry, and relates to the technical field of microbiological detection.The primer probe composition is composed of a multiple PCR primer composition and a single-base extension probe set and can conduct PCR amplification and extension on conservative and stable areas in the multiple cattle and sheep diarrhea pathogens; the extension product suitable for MALDI-TOF MS detection is obtained. The multiplex PCR primer composition, the single-base extension probe set, the kit and the method show good specificity and sensitivity, and show excellent detection performance. Meanwhile, the detection flux and the detection efficiency are obviously superior to those in the prior art, and the primer has obvious application potential in rapid detection of the nucleic acid of the important diarrhea pathogens of cattle and sheep.

The invention belongs to the technical field of gene detection, and relates to a primer group and a kit for multiple PCR (Polymerase Chain Reaction) detection of a bovine diarrhea virus. The SapV, BNoV and BEV triple PCR detection technology established in the invention has good specificity and repeatability, can be used for rapid detection of cattle farm diarrhea virus mixed infection, and lays a technical foundation for bovine viral diarrhea pathogen epidemiological investigation.

The invention provides a synchronous detection primer for type II norovirus, group A rotavirus and adenovirus, which comprises specific primer pairs designed for conserved genes of three pathogens of type II norovirus, group A rotavirus and adenovirus, namely an ORF1 / ORF2 primer pair, a VP6 gene primer pair and a Hexon gene primer pair. The invention also provides a detection probe, which comprises an ORF1 / ORF2 capture probe, a VP6 capture probe and a Hexon capture probe. Meanwhile, the invention further provides application of the primer and the probe in preparation of infectious diarrhea related virus detection products, a specific detection kit and a specific detection method. According to the present invention, the multiple detection of three common diarrhea viruses can be achieved, the detection can be completed in a short time, the amplification product can be rapidly and accurately identified, the color development can be performed, the detection sensitivity and the specificity can be improved, the large-scale instrument is not required, and the cost is low.

The invention relates to the technical field of molecular biology and virology, in particular to an LAMP (loop-mediated isothermal amplification) primer composition for detecting porcine epidemic diarrhea virus and application thereof, the LAMP primer composition comprises an external primer pair, an internal primer pair and a loop primer pair; wherein the external primer pair comprises a forward primer F3 and a reverse primer B3; the internal primer pair comprises a forward primer FIP and a reverse primer BIP; the loop primer is a loop reverse primer LB. The LAMP primer composition for detecting the PEDV has the advantages of high sensitivity and high specificity, the PEDV can be rapidly and efficiently detected, and the operation environment does not need to be strictly controlled. The kit prepared from the LAMP primer composition has the advantages of high efficiency, reliability, simplicity and rapidness in operation, convenience in carrying and low detection cost, and can be prospectively used for early diagnosis of PEDV (porcine epidemic diarrhea virus).

This invention belongs to the field of virus detection technology, specifically relating to a monoclonal antibody against porcine delta coronavirus N protein and a latex immunochromatographic test strip containing this antibody. This invention uses prokaryotically expressed PDCoV N protein as an immunogen to immunize mice, and screens for specific monoclonal antibodies 2G3 and 9C4 through cell fusion and subcloning techniques. The test strip includes a PVC base plate, a sample pad, a conjugation pad, a nitrocellulose membrane (NC membrane), and an absorbent pad; a control line C and a detection line T are respectively set on the NC membrane. This test strip can specifically recognize PDCoV, with a detection limit as low as 10. 3.0 TCID 50 The virus has a concentration of 1000 μL / mL and does not cross-react with other common clinical diarrhea viruses. The results can be interpreted in just 15 minutes, making it fast, sensitive, and easy to operate.

The invention provides a primer probe combination for detecting diarrhea viruses through quadruple TaqMan qPCR and application of the primer probe combination, and belongs to the technical field of diarrhea virus detection. The primer probe combination for detecting the diarrhea virus through quadruple TaqMan qPCR provided by the invention comprises primer probes for detecting rotavirus, norovirus, human astrovirus and adenovirus respectively. The primer probe combination provided by the invention has the advantages of high sensitivity, stability, specificity and the like when being used for quadruple TaqMan fluorescent quantitative PCR detection.