56 results about "Serum neutralization" patented technology

The invention discloses an immune-electrochemical sensor based on an AuNPs@AgNCs nano composite material, construction and applications thereof. Ab1 is fixed by graphene, PSA is taken as the target analyte, SA-AuNPs@AgNCs labeled by streptavidin is easily combined with monoclonal antibody Ab2(biotin-Ab2) which has been modified by biotin, and thus a PSA immunity sensor can be prepared by a sandwich interlaying method through the specific reactions between an antibody and an antigen. When the immune sensor is used, silver and H2O2 are precipitated on the sensor through electric catalytic reduction reactions, the detection is rapid and sensitive, has good specificity, and is capable of detecting the concentration of PSA with an ultralow content. The concentrations of f-PSA and t-PSA in serum of prostatic hyperplasia patients and prostatic cancer patients can also be detected by the sensor, so the sensor has an important meaning for diagnosis and identification on gray areas in prostatic cancer diagnosis.

The invention relates to preparation of a vaccine for hand-foot-and-mouth disease viruses and an application method thereof, belonging to a novel vaccine for preventing infectious diseases. The vaccine of the invention mainly comprises the components of high-purified inactivated human enteropathogenic virus 71 (EV 71) and an aluminum adjuvant. The vaccine, which is prepared according to the method of the invention, has excellent immunogenicity, and after immunity, organisms can selectively generate a high titer serum neutralization antibody, thereby preventing infectious diseases caused by the human EV 71.

The invention discloses a preparation method and a special culture medium of veterinary D-type clostridium perfringen toxin. The culture medium per 100ml comprises the following components: 1-1.5g soy peptone, 1-1.5g casein peptone, 0.5-0.75g yeast extract, 0.5-0.75g Na2HPO4.12H2O, 1-1.5g dextrin and the balance of water, wherein a pH (potential of hydrogen) value of the culture medium is 8.0-8.5. The preparation method of the D-type clostridium perfringen toxin comprises the steps of inoculating a D-type clostridium perfringen production strain into the culture medium, collecting a culture material, performing centrifugation, and then filtering a supernate. With the adoption of the method, the maximum toxicity can be improved to be 45 times a seedling standard in Regulations on National Veterinary Biological Products, and an output-input ratio can be increased to be 30-225 times that of the original traditional technology. In addition, corresponding serum neutralization titer of a toxoid vaccine prepared by the D-type clostridium perfringen toxin on a rabbit and a sheep is also improved to be 8.3 and 13.3 times a regulation standard respectively.

The invention discloses a detecting kit for hepatitis a virus IgM and IgG antibodies through a colloidal gold method. The detecting kit comprises a kit body, sample diluting liquid and detecting test paper. The detecting test paper is composed of a plastic supporting plate, a sample feeding pad, a colloidal gold pad of antigen VP1 marked with hepatitis a virus specifity, a nitrocellulose NC film, a detecting line T1, a detecting line T2, a quality control C line and a sample absorbing pad and arranged in the kit body. The kit detects that IgM and IgG in serum of the sample are positive simultaneously, the sample result is completely identical with an enzyme-linked immunosorbent assay (ELISA) reagent, no remarkable difference exists, and the kit is applicable to clinical detecting. The kit is low in cost and simple in preparation process and has practical value.

The invention provides a preparation method of an oral inactivated porcine epidemic diarrhea virus microcapsule and relates to the field of preparation of microcapsules. The invention aims at overcoming the technical defects in an oral inactivated vaccine in domestic porcine epidemic diarrhea virus prevention and control at the present stage and provides a preparation method for the oral inactivated porcine epidemic diarrhea virus microcapsule based on inactive porcine epidemic diarrhea virus and oral microcapsule preparation technologies. Animal experimental results find that a serum neutralizing antibody titer detection result shows that the microcapsule vaccine can induce piglet mucosal immunization and systemic immune response, and serum and mucosa antibodies with relatively high levelare produced, so that the microcapsule vaccine is objectively explained to have a certain slow-release effect. After a microcapsule vaccine antigen substance is orally taken by piglets, expression ofspecific antibodies IgG and IgA of the piglets can be induced, resistance of the piglets on PEDV is improved, death rate of the piglets is reduced, and toxicity attack protection rate is 60-75% higher than that of a control group.

The present invention discloses a scutellaria barbata polysaccharide antitumor mechanism detection research method which comprises the following detecting steps: an in-vivo tumor suppression experiment; ANAE method determination of the total number of T cells in peripheral blood of tumor-bearing mice; flow cytometry determination of T cell subsets and T cell surface protein-CD28 in the peripheral blood of the tumor-bearing mice; and determination of content of IL-2 and IFN-gamma in serum of the peripheral blood of the tumor-bearing mice; the scutellaria barbata polysaccharide in-vivo antitumor and cell immune function influence detection research method discloses a major role of scutellaria barbata polysaccharide in tumor treatment, and meanwhile the study on body cell immune mechanism of plant polysaccharides can be promoted.

The invention provides a method for simultaneously detecting serum 24,25(OH)2D and 25OHD by liquid chromatography tandem mass spectrometry. The method comprises the following steps: 1) preparing a serum sample; 2) separating the sample through liquid chromatogram, and removing an interference substance; 3) obtaining a detection signal through mass spectrometry; 4) drafting a standard curve; and 5)detecting the signal intensity of 24,25(OH)2D3, 24,25(OH)2D2, 25OHD2, and 25OHD3 in the to-be-measured sample, substituting the signal intensity in the above standard curve for calculation, and realizing the quantitative determination of 24,25(OH)2D and 25OHD in serum. The rapid detection method for simultaneously detecting 24,25(OH)2D and 25OHD by using less sample (200 [mu]l of serum) based onliquid chromatography tandem mass spectrometry, and can distinguish the interference of 3-epi 24,25(OH)2D3and 3-epi 25(OH)D3.

The invention belongs to the technical field of medical biology, and discloses a determination kit for simultaneously detecting contents of aldehyde ketoreductase 1B10 and alpha fetoprotein by using atime-resolved fluorescence immunoassay method, which can be used for screening, diagnosing, curative effect judging, prognosis evaluating or recurrence monitoring of diseases such as liver cancer, hepatitis, liver cirrhosis and the like. The kit comprises an elisa plate, a calibrator, a europium-labeled AKR1B10 detection antibody, a terbium-labeled AFP detection antibody and an enhancement solution. The AKR1B10 and the AFP in the serum to be detected are respectively combined with the AKR1B10 and the AFP coated antibody of the elisa plate, then two detection antibodies are added to form a double-antibody sandwich compound, an enhancement solution is added to dissociate europium ions and terbium ions on the compound, and high-intensity fluorescence is respectively emitted at wavelengths of340nm and 295nm. The fluorescence intensity is in direct proportion to the concentrations of AKR1B10 and AFP in the sample. By adopting the kit, the contents of the two markers can be detected at thesame time, and the detection sensitivity and specificity are greatly improved.

The invention discloses a clostridium perfringens type B exotoxin, a preparation method thereof, a toxigenic culture medium and an application thereof. Each 1000 ml of the toxigenic culture medium isprepared from the following raw materials by weight: 15-25 g of peptone, 3-5 g of yeast extract, 3-4 g of NaCl, 5-10 g of Na2HPO4.12H2O, 5-10 g of dextrin or 5-10 g of coarse dextrin, and the balanceof water for injection. The virulence of the clostridium perfringens toxin liquid produced by the method according to the present invention is up to 1 mouse MLD of no more than 0.0006 mL, and the virulence is 3.3 times than the vaccine production standard. Moreover, a triple inactivated vaccine against bradsot, struck (or lamb dysentery) and enterotoxemia prepared by the clostridium perfringens type B exotoxin has serum neutralizing titer in rabbits and sheep equal to or higher than the relevant standard.

The invention relates to the technical field of gene engineering, in particular to a recombinant gene of a rabies virus, a recombinant pseudovirus and a construction method and application of the recombinant pseudovirus. The nucleotide sequence of the recombinant gene is as shown in SEQ ID NO: 1. The preparation method comprises the following steps: replacing VSV-G in a lentivirus envelope plasmid with a recombinant gene RABV-G-N to construct a recombinant envelope plasmid pCMV-RABV-G-N, and then co-transfecting a human embryonic kidney cell HEK-293T with the recombinant envelope plasmid, pLV-eGFP containing a green fluorescent protein reporter gene and psPAX2 to obtain the recombinant pseudovirus. The recombinant gene provided by the invention has biological characteristics of natural rabies virus, and can improve pseudovirus immunogenicity, and the prepared recombinant pseudovirus has high packaging titer, and can replace natural rabies virus to evaluate serum neutralizing antibody titer.

The invention provides a vaccine strain rSN-R92G-E93K, and belongs to the technical field of influenza vaccine preparation. The construction method comprises: preparing an expression vector pHW-SN-HAfrom a HA gene fragment; carrying out point mutation on the expression vector; co-transforming cells by using the obtained expression plasmid SNHA-R92G-E93K and plasmids containing other genes; and carrying out recombination. According to the present invention, the vaccine strain rSN-R92G-E93K has the HI titer of 11.6+/-0.5 log2 and the serum neutralizing titer of 296.50+/-103.95 log2, such that the vaccine strain rSN-R92G-E93K is an ideal vaccine strain.

The invention discloses a bovine viral diarrhea virus low virulent strain and application thereof.The low virulent strain is obtained after 100 generations of continuous passage culture of an SMU-Z6/1a/SC/2016 strain, the nucleotide sequence of the low virulent strain is shown as SEQ ID NO.1, and the microbial preservation number of the low virulent strain is V202131. After BABL/c mice are immunized with the bovine viral diarrhea virus attenuated strain, high serum neutralizing antibody, serum IgG antibody, lung and intestinal sIgA antibody and splenic lymphocyte proliferation rate can be generated, and a good immune effect is achieved. The bovine viral diarrhea virus attenuated strain has a very good application prospect in prevention, treatment and diagnosis of the bovine viral diarrhea virus.

The invention relates to an adaptive culture method of a rotavirus P(2) G3 strain and a P(8) G1 strain on KMB17 cells and immunogenicity, belonging to the field of virus culture methods. 10 generations of adaptive culture of the two strains of rotavirus are continuously carried out on the KMB cells, thereby gradually increasing the cytopathy along with the virus generation, gradually increasing the infective titers and stabilizing the infective titers at the higher level. The virus titers respectively achieve 7.75CCID50/ml and 7.33CCID50/ml. The virus is applicable to the KMB17 cells and has a stable genome, and the neutralizing antibody titers in serum of mice immunized with the copied virus are 1:16384 and 1:8192 respectively. The rotavirus P(2) G3 strain and the P(8) G1 strain formed by the adaptive culture can be stabilized copied in the KMB17 cells, and the produced virus can keep the basic biological property of parent virus strains and have better immunogenicity.

The invention discloses a preparation method of rabbit anti-canine parvovirus 2a type polyclonal antibodies. The method comprises the following steps of: A1) performing virus amplification; A2) concentrating antigens; A3) measuring the content of antigens; A4) adopting the immunization method; and A5) purifying serum; purifying the canine parvovirus (CPV)-2a virus strain, amplifying, concentrating the obtained virus liquid with polyethylene glycol (PEG)6000, emulsifying with Freud's adjuvant, and performing subcutaneous immunizations at ten different points, wherein basic immunization and booster immunization are totally performed four times. The prepared polyclonal antibodies have high titer, the hemagglutinin inhibition (HI) value is no less than 1:10240, and the serum neutralization (SN) value is no less than 1:51200; and compared with the CPV-2 polyclonal antibodies, the neutralizing CPV-2a antibodies have a stronger effect.

The invention discloses a duck liver positive serum preparation method, and belongs to the technical field of virus detection. The duck liver positive serum preparation method comprises the steps that Step 1, materials are prepared, and a virus strain, an inactivated vaccine and a chick embryo are optimized; Step 2, immune serum is produced in a trail manner, the inactivated vaccine is injected into the chick embryo, so that the chick embryo generates immune reaction, and finally, the chick embryo is promoted to generate the immune serum; Step 3, a safety test and an effect test are performed on the chick embryo by using the immune serum; Step 4, the immune serum is prepared on a large scale; Step 5, product quality inspection and safety inspection are performed on the prepared immune serum; and Step 6, the immune serum is efficiently stored. According to the invention, the neutralizing reaction with the type 3 duck hepatitis virus is poor, the titer of the neutralizing antibody is 1:2896, the titer of the neutralizing antibody is stable after the repeated test, the antibody can be used for inspection, and during neutralization of the virus liquid with different contents by using the serum with different neutralizing antibody titers, and the virus liquid with the concentration of 104.0 ELD50/0.1 ml can be completely neutralized when the titer of the neutralizing antibody is 1:2048 according to the result.

The invention relates to the technical field of biomedicine, in particular to a DNA methylation-based invasive glioma classification device, which comprises a detection module, a classification module and an information display module, whether neuroglial cells have canceration or not and the canceration degree are predicted by measuring the methylation degree of a CpG island, neuroglioma is classified, and by adopting real-time quantitative methylation specific PCR, the sensitivity and the accuracy are high, the DNA methylation condition of formalin soaked specimen tissue can be detected, and the detection result is accurate. Therefore, the real-time quantitative methylation specific PCR has the potential of being used for glioma diagnosis, mi RNA can stably exist in serum and is easy to obtain and has the potential of becoming an early diagnosis molecular marker, and the content of mi R-25 and mi R-223 in the serum can be used as a diagnosis marker of a non-small cell lung cancer patient; a specific glioma mi RNA marker is screened out, DNA is extracted from blood of a patient, then the mi RNA expression level can be detected through PCR, and therefore early diagnosis can be conducted on glioma.