60 results about "Artificial infection" patented technology

The invention provides a multi-antigen enzyme linked immunosorbent assay (ELISA) kit for detecting an African swine fever virus (ASFV) antibody, belonging to the field of a biotechnology and diagnosis and research of animal-borne diseases. The multi-antigen enzyme linked immunosorbent assay kit comprises expression and purification of three types of ASFV recombined antigens, preparation of positive and negative control blood serum of an ASFV antibody, optimal envelope antigen combination and concentration determination, optimization of multi-antigen ELISA (MA-ELISA (Microalbumin-Enzyme Linked Immunosorbent Assay)) reaction parameters, determination of an ASFV antibody negative blood serum critical value, MA-ELISA detection artificial infection and determination of sensitivity, specificity and repeatability of field blood serum samples. By detecting and testifying a large quantity of known blood serum samples, the sensitivity, the specificity and the repeatability of detecting the ASFV antibody by the MA-ELISA are obviously higher than those of an ELISA method recommended by World Organization for Animal Health and oversea similar kits; and the multi-antigen enzyme linked immunosorbent assay kit can be used for ASFV serological diagnosis, epidemiological investigation and live pig import and export quarantine inspection.

The invention discloses hyper-immune serum prepared by immunizing a hybridized pig with the age of 50-60 days by taking an inactivated vaccine of porcine epidemic diarrhea virus ZJ08 strain CGMCC No.7806 as an immunogen due to immunization for three times. The prepared hyper-immune serum is high in properties, high in specificity and high in safety and is not polluted by bacteria, moulds, mycoplasma and exogenous viruses, and the artificial infection cure rate is 100 percent. The neutralization titer of the serum is over 26, the serum is separately filled at the temperature of minus 70 DEG C, and the retention period is 12 months.

The invention relates to a method for the fish genealogy establishment and the disease resistance and high yield improved type selection. By adopting the technologies, such as group selection, genealogy selection, hybridization and artificial infection, as well as the sieving of the quick growth genealogy and disease resistance genealogy, the genealogy which has the advantages of strong flounder disease resistance and quick growth is selected, and the selection method for the disease resistance and high yield high quality improved type fish is established. The technology has the advantages of strong pertinence, high selection efficiency, obvious effect, etc. and the survival percent of the disease resistance flounder fish fry is more than 80 percent, which is increased by 30 percent more than the survival percent of the common flounder fish fry; the growth speed of the disease resistance flounder fish fry is also increased by more than 10 percent. The method in the invention opens up a new technical approach for culturing the disease resistance fish variety. As a disease resistance high yielding variety selection method applied to the improved fish type selection, the method is suitable for the popularization and the application on all the cultured fishes, and has important significance and application value in culturing the cultured fish of disease resistance and high yield improved type.

The invention relates to a method for constructing tongue sole families and breeding good families, which comprises the technical content of breeding a tongue sole group, constructing the tongue sole families and screening a disease-resistant family and a fast growing family of tongue sole. The method comprises the following steps of: constructing the families through the group breeding and intraspecific hybridization technology; then, screening out the family resisting vibrio anguillarum disease from a large amount of family by an artificial infection approach, and screening out the fast growing family by growth comparison. The fries in the good families can be directly popularized and applied, or used for breeding a high-yield and disease-resistant good strain of tongue sole. The invention has the characteristics of strong pertinence, high breeding efficiency, obvious effect and the like. The established method for screening good tongue sole families provides a new technical approach for breeding the high-yield and disease-resistant good strain of tongue sole. The method is suitably popularized and applied to other cultured fishes, and has important significance and application value for breeding high-yield and disease-resistant strains of the cultured fishes.

The invention aims to provide a brucellosis antibody competitive enzyme-linked immunosorbent assay reagent kit which meets the demand of epidemic prevention of animals in our country, is sensitive and specific, and can accurately diagnose brucellosis. The reagent kit is assembled by utilizing lipopolysaccharide of smooth brucella as an antigen coated ELIAS plate, using an inactivated bacterial liquid to immunize a healthy cow or artificially infect the healthy cow to prepare positive control serum and standard weak-positive serum, using serum of a healthy non-immune cow as negative control serum, using a monoclonal antibody as a competitive antibody, namely an ELIAS secondary antibody (goat anti-rat IgG), and preparing a serum diluent, a washing liquid, a substrate solution A, a substrate solution B, H2O2, and a stop buffer. The brucellosis antibody competitive enzyme-linked immunosorbent assay reagent kit is a competitive ELISA reagent kit which is sensitive and specific and can accurately diagnose the brucellosis. The reagent kit meets the demand of the epidemic prevention of the animals in our country, and performs final definite diagnosis on the brucellosis by means of laboratory diagnosis technology.

The invention discloses an egg yolk antibody for preventing and treating porcine epidemic diarrhea and a preparation method of the egg yolk antibody. The preparation method comprises the following steps: using an inactivated vaccine of a porcine epidemic diarrhea virus as immunogen for immunizing a laying hen; harvesting egg yolk liquid and purifying from the egg yolk liquid to obtain the egg yolk antibody, wherein a porcine epidemic diarrhea virus strain is PEDV/CH/2014, with a preservation number of CGMCC (China General Microbiological Culture Collection Center) No. 10111. The egg yolk antibody prepared by the preparation method disclosed by the invention has the advantages of good characters, easiness in storage, high safety and good preventing and treating effects; by an in vitro neutralization test, the neutralizing titer of the egg yolk antibody is measured to reach 1:128; by means of a clinical prevention test, the diarrhea incidence of tested pigs can be reduced by 20 percent, and the cure rate of artificial infection reaches 100 percent.

The invention aims to provide a new-generation brucellosis antibody competition enzyme-linked immunosorbent adsorption test detection kit which meets animal epidemic prevention requirements in China, is sensitive and specific and can accurately diagnose brucellosis. In the invention, the lipopolysaccharide of smooth type brucella is used as an antigen-coated enzyme-labeled plate, inactivated bacilli liquid is used for immunizing or artificially infecting a healthy cattle to prepare positive control sera and standard weak positive sera, healthy non-immune cattle sera are used as negative control sera, a novel monoclonal antibody is used as a competition antibody, the protein AG enzyme-labeled substance of an anti-Fc receptor is used as an assistant antibody, serum diluent, a washing solution, a substrate solution A, a substrate solution B, H2O2 and a stop solution are prepared, and all raw materials are assembled to form the kit. The invention is a competition ELISA kit which is sensitive and specific and can accurately diagnose brucellosis. The invention meets animal epidemic prevention requirements in China, and carries out last diagnosis on the brucellosis by means of a laboratory diagnosis technique.

The invention discloses an antagonistic drug against replication of a porcine reproductive and respiratory syndrome virus (PRRSV) and application of the antagonistic drug. A virus infection test is adopted, and Xanthohumol (Xn), the antagonistic drug of the PRRSV, is discovered for the first time from a natural medicine library with 386 plant sources. The drug can effectively inhibit PRRSV replication in both Marc-145 and PAM cells, five xanthohumol derivatives with different molecular structures are artificially synthesized, and the viral infection test further finds that the inhibition effect to virus replication by drug derivative Xn-4 is highest in vitro; and test results of artificial infection and drug therapy in piglets show that the Xn-4 can effectively inhibit PRRSV viremia, relieves clinical symptoms of infected pigs, and significantly reduces pulmonary inflammation and pathological damage. Combined with the results, it is proved that the antagonistic drug has important preventive and therapeutic effects to a porcine reproductive and respiratory syndrome, and has important application prospects.

The invention relates to a method for culturing cordyceps sinensis by performing endogenous pressurization and artificial infection of hirsutella sinensis on hepialus larvae, belonging to the technical field of artificial culture of the cordyceps sinensis. The method comprises the steps of: mixing the following components in percentage by weight: 28-34% of bombyx mori pupae, 5-7% of potatoes, 0.008-0.01% of magnesium sulfate, 0.035-0.05% of potassium dihydrogen phosphate and the balance of water, boiling, filtering and sterilizing to prepare an aseptic infection stock solution; taking the hirsutella sinensis as an infection bacteria source, picking pure mycelia through inoculating needles, placing the pure mycelia into the aseptic infection stock solution according to the proportion that the hirsutella sinensis picked by 10-15 needles are placed into every 5ml of the aseptic infection stock solution, grinding and then preparing an infection source bacterial solution; and sucking the infection source bacterial solution through a syringe, inserting a needle head from the excretory opening of each hepialus larva to press 0.3-0.5ml of the infection source bacterial solution into the intestinal tract of the larva, then placing the larvae in the shade and culturing under the conditions that the temperature is 17-21 DEG C and the relative humidity is 78-85% for more than 15 days. The method disclosed by the invention has the advantages of infection rate of up to above 90 percent, short period, low cost and great application prospects for large-scale culture of the cordyceps sinensis.

The invention which belongs to the technical field of biological detection relates to a method for detecting MP of sheep and provides the methylation state of CpG islands in an MBL (mannan-binding lectin) gene promoter region related with the MP of sheep and an application thereof. The invention also provides a method for detecting the MP of sheep and concretely relates to the methylation detection of the MBL promoter region and the artificial infection verification. The methylation state detection of the MBL gene promoter region of sheep is carried out through selecting a bisulfite sequencing process, the methylation state and the frequency of each CpG locus in a given region can be accurately detected, three substantially differential CpG methylation loci, CpG3, CpG4 and CpG5, related with the MBL level are found, a case that the MBL level after toxic elimination is lower than the MBL level before the toxic elimination is promoted, and the MBL gene promoter region methylation may be a more substantial characteristic of the MBL level reduction of serum. The method allows the MP infection of sheep and susceptible individuals to be sensitively and accurately detected, and provides complete information and scientific bases for the sheep MBL gene promoter region methylation and the MP infection research of sheep.

The invention belongs to the field of aquatic product disease prevention and treatment, and relates to a method for building a gyrodactylus purebred artificial infection system, in particular to a method for building an artificial infection system of gyrodactylus kobayashii hukuda purebred parasitized on the gill and the body surface of goldfish. The method includes the steps of (1) obtaining healthy and gyrodactylus-free goldfish; (2) conducting gyrodactylus artificial infections on the healthy and gyrodactylus-free goldfish; (3) maintaining the state after the gyrodactylus artificial infections are carried out. By means of the method for building the gyrodactylus purebred artificial infection system, an animal model is provided for screening medicine, developing new fish medicine and the like with the goldfish as hosts.

The invention relates to a method for determining the activity of a trichophyton rubrum keratinase. The method comprises the following steps: (1), constructing an artificial skin containing epidermis and dermis structures: resuspending human skin fibroblast cells in a collagen liquid, dropwise adding into a cell culture well plate and culturing; after the cells are completely expanded, dropwise adding a human keratinocyte suspension to the surface of collagen gel and continuously culturing; carrying out air-liquid interface culture when the human keratinocytes completely cover the surface of the collagen gel; and stopping culturing until a cuticular layer is differentiated into 5-8 layers; (2) carrying out artificial infection on trichophyton rubrum; and (3) determining the activity of the keratinase. The artificial skin, which is constructed by the method disclosed by the invention, becomes a natural inducer for production of the keratinase and has the following advantages that firstly, the artificial skin contains the dermal layer, the epidermal layer and 5-8 cuticular layers and the structure and function are closer to those of the human skin; secondly, keratin secreted by the keratinocyte in the epidermal layer is used as a natural substrate to induce the production of the keratinase; and, thirdly, the keratinase produced by inducing the natural substrate keratin, is accurate, real and stable in the determination of the activity.

The invention relates to a preparation method and use of a duck-derived coronavirus attenuated strain DCV35. The preparation method is characterized in that a duck-derived coronavirus virulent strain IBV ZZ2004 (with microbial preservation number of CGMCC NO.3842) separated by the inventor is subjected to passage attenuation, the S1 gene sequence of the screened attenuated strain DCV35 has obvious characteristics, and the variant sequence has stable heredity in 22th-35th generations. In SPF chicken infection adopting an artificial infection test and a horizontal transmission test, the screened attenuated strain DCV35 does not produce virulence enhancement. An experiment proves that after inoculation of a chicken with the attenuated strain, the attenuated strain can effectively activate a chicken immunization system, has good effects of preventing chicken infectious bronchitis and can be used as a vaccine for preventing chicken infectious bronchitis.

The invention relates to an artificial infection method of a tree coral. The technical scheme is described as follows: firstly, randomly dividing strains obtained and separated from a diseased coral into a plurality of test groups; simultaneously carrying out artificial infection on healthy corals by utilizing the parallel groups of mixed bacterium liquid; respectively carrying out artificial infection on the healthy corals by utilizing the strains of the test groups capable of inducing the healthy corals to be diseased; and finally, confirming the bacterium pathogens capable of inducing the healthy corals to be diseased, therefore, processes of rapidly and accurately separating and identifying the coral bacterium pathogens are realized. The method disclosed by the invention has the beneficial effects that: aiming at coral bacterium diseases, the rapid separation and confirmation of pathogenic bacteria are realized; and compared with a traditional method, the method is capable of greatly reducing the working intensity of coral bacterial disease epidemiological investigation.

An equivalent and quantitative test method of white spot syndrome virus resistance of prawns belongs to the technical field of disease-resistant prawn variety selection and includes: firstly, preparing poison bait; secondly, testing WSSV (white spot syndrome virus) resistance; thirdly, acquiring data and varieties; and fourthly, determining reserved parent seeds. The type of infecting virus is determined and the number of virus copies is detected by TaqMan real-time fluorescent quantitative PCR (polymerase chain reaction), the prawns having the same number of virus copies are used to prepare the poison bait, and the number of virus copies in muscular tissues in specific weight is calculated according to actual artificial infection needs. The equivalent and quantitative test method of the prawns is achieved by adding edible red pigment in the poison bait and infecting the prawns to be tested one by one, and obtaining true and reliable data is guaranteed.

The invention relates to the technical field of livestock and poultry pathogenesis model establishment, in particular to an establishing method of a chicken H9 subtype avian influenza artificial infection model. In the method, newly-developed SPF hens are used as test animals, the environment temperature is controlled to be 10-15 DEG C in the whole test period two days before infection, H9 subtypeAIV and poultry source escherichia coli virulent strains and IBV M41 strains are adopted for sequential co-infection through a nasal inhalation mode, the infected hens show obvious fevering, droopingspirit and cough and asthma symptoms, autopsied diseased hens have characteristic lesions of the H9 subtype avian influenza, and the model is stable and reliable. By means of the method, the obtaineddisease model is stable and typical, and the important basis is laid for subsequently carrying out the avian influenza medicine prevention and cure research.

The invention discloses a feed additive enteric-coated sustained-release pellet with a function of preventing and treating poultry respiratory syndromes and a preparation method for the feed additive enteric-coated sustained-release pellet. The enteric-coated sustained-release pellet is prepared from the following raw materials: water-soluble vitamin A, water-soluble vitamin D, water-soluble vitamin E, thymol, cinnamyl aldehyde, menthol, eucalyptus oil, maltodextrin, sodium carboxymethylcellulose, lactose and a coating solution which is a Eudragit enteric-coated coating solution with a model of Eudragit L30D-55, wherein the coating solution is water suspension with mass concentration of 5%-8%. The feed additive enteric-coated sustained-release pellet with the function of preventing and treating poultry respiratory syndromes disclosed by the invention has a treatment effect on sick chicken with respiratory syndromes due to artificial infection of chicken infectious bronchitis virus (M41 strain), can effectively improve the respiratory tract function, is small in dosage, quick in effect-taking, and has the effects of promoting growth and increasing a feed conversion rate.

The invention discloses a traditional Chinese medicine composition for preventing a pullorum disease. The traditional Chinese medicine composition comprises the following medicinal materials in parts by weight: 120 to 180 parts of red-knees herb, 120 to 180 parts of cablin potchouli herb, 120 to 180 parts of glossy privet fruit and 80 to 120 parts of astragalus. A test shows that the resistance of a chick to artificial infection of pullorum disease salmonella can be obviously improved, a strong sterilizing effect is expressed, the weight gain rate of the chick can be kept in a normal level, and the survival rate of the chick infected with the pullorum disease salmonella can be improved. Moreover, the traditional Chinese medicine composition has the characteristics of safety, non-toxicity and no drug residue, and the quality of a chicken product can be improved. The traditional Chinese medicine composition can replace antibiotics and feed additives for the poultry, the pollution of livestock products caused by drug resistance and drug residue can be effectively avoided, harmful livestock excrements, protein dosage and nitrogen pollution are reduced, and the market prospect is wide. Meanwhile, a preparation method is simple, convenient to use and lower in cost.

The invention relates to a novel use of epigallocatechin gallate, namely an application of the epigallocatechin gallate in preparation of a medicine for prawn white spot syndrome control. The application is advantageous in that tests prove that the epigallocatechin gallate (EGCG) can antagonize invasion infection of WSSV viruses and can control the prawn white spot syndrome. Artificial infection tests prove that the death rate of prawns infected with the WSSV viruses shows a decreasing trend along with increases of the EGCG concentration, and the death rate of prawns when the EGCG concentration during toxic material elimination is 20 [mu]g/mL and 40 [mu]g/mL is only about 20%.