67results about How to "Reduce Biosecurity Risks" patented technology

The invention relates to a bovine brucella indirect ELISA antibody detection kit. The kit uses the lipopolysaccharide (LPS) of brucella suis S2 acclimated and induced by the application institution in the 1960s as the coating antigen, and improves the sensitivity of detection. The study improves the LPS purification and purity determination method, and endows the developed kit with good specificity. The kit not only can effectively eliminate the interference of conventional Gram negative bacteria to Brucella, but also can get rid of the technical defect that general brucellosis indirect ELISA (enzyme-linked immunosorbent assay) kits cannot distinguish enterocolitis Yersinia O9 and Escherichia coli O157 interference, and improves the specificity of detection. Through improvement and optimization of the main reagent formula, the reaction background of the kit is effectively controlled, and also the time required by detection is shortened.

The present invention relates to a rapid, efficient and accurate bovine Brucella colloidal gold diagnostic method. According to the present invention, Brucella strain S2 lipopolysaccharide (LPS) is adopted as a testing line coating antigen so as to improve the detection sensitivity; the LPS purification and purity determination method is improved, such that the developed kit has good specificity; the monoclonal antibody on the Fc terminal of the mouse anti-bovine IgG is used to label the colloidal gold to prepare the colloidal gold pad of the colloidal gold antibody detection test paper strip so as to further improve the detection specificity; the bovine Brucella colloidal gold antibody detection test paper strip can be used for detecting the Brucella antibody in the bovine serum, and has characteristics of strong specificity, high sensitivity, convenience, rapidness, and the like; and the complex detection equipment is not required for the Brucella detection, and the bovine Brucella colloidal gold antibody detection test paper strip is especially suitable for cattle farm breeding staffs and grassroots veterinarians.

The invention discloses silver-carrying antibacterial hydrocolloid dressing and a preparation method thereof. The silver-carrying antibacterial hydrocolloid dressing is sequentially provided with a backing, a hydrocolloid functional dressing core and an isolation layer from top to bottom; the hydrocolloid functional dressing core is prepared from the following raw materials including sodium carboxymethylcellulose, an SIS thermoplastic elastomer, liquid paraffin, tackifying resin, an antioxidant, dioctyl adipate as a plasticizer, a bacteriostatic agent and a collaborative bacteriostatic agent. In a preparation process, the silver ion bacteriostatic agent and the plant extract collaborative bacteriostatic agent are added; with addition of the plant extract collaborative bacteriostatic agent, the addition concentration of the silver ion bacteriostatic agent is lowered when the antibacterial activity of the dressing is effectively improved; with addition of the silver ion bacteriostatic agent, the antibacterial wide adaptability and the antibacterial durability are both improved; and as the two bacteriostatic agents are used in a matching manner, a wound can be effectively prevented from being infected, and the silver-carrying antibacterial hydrocolloid dressing also has the advantages of blocking bacteria, absorbing exudate, avoiding wound adhesion, supplying an ideal wet healing environment for a wound, promoting healing of the wound and reducing formation of scars.

The invention relates to a SARS-CoV-2 inactivated vaccine and a preparation method of the vaccine. The preparation method comprises the following steps: step (1) recovery and large-scale culture of Vero cells; step (2) inoculation of working seeds, batch of virus seeds, virus culture and one-time harvest virus liquid, namely, the virus harvest liquid; step (3) obtaining an inactivated virus concentrated solution after a first virus inactivation, concentration and a second virus inactivation; step (4) obtaining a vaccine stock solution after purifying the inactivated virus concentrated solution; and step (5) preparation of semi-finished product and sub-package after the stock solution is verified to be qualified.

The invention relates to a colloidal gold test paper bar for antibody detection of sheep Brucella. In the invention, lipopolysaccharide (LPS) extracted from a Brucella vaccine strain (S2) is taken as an envelope antigen of the colloidal gold test paper bar and improves the sensitivity of detection. Monoclonal-antibody-labeled colloidal gold of a mouse anti-sheep IgGFc terminal is prepared into a gold colloid pad of the colloidal gold test paper bar for antibody detection, and nonspecific adsorption of antibody protein by SPA in the tradition is changed. The monoclonal-antibody-labeled colloidal gold is taken as a colloidal gold labeled protein, so that specificity of detection is further improved. A set of serum for controlling quality of the test paper bar is prepared, and specificity and sensitivity of the test paper bar is ensured. The test paper bar can be used for on-site sampling, is convenient and rapid to use, and is quite suitable for application to a basic farm or a basic veterinarian.

The invention relates to a clostridium perfringens Beta toxin recombination subunit vaccine and a production method thereof. The prepared clostridium perfringens Beta toxin recombination subunit vaccine is produced by using a recombination clostridium perfringens Beta toxin protein which is processed by codon optimization and contains 4 amino acid mutations, namely integrity and spatial conformation of a natural toxin protein are reserved in the greatest degree, so immunogenicity of the natural toxin protein is kept, and a biological potential safety hazard caused by the single amino acid mutation is avoided. The vaccine further has the advantages of simple preparation process, low immunizing dose, good vaccine efficacy and the like. Compared with a current commercial clostridium perfringens natural toxin inactivated vaccine in China, a bio-safety risk in a vaccine production process is greatly reduced. The vaccine is a perfect candidate vaccine for updating a current C-type clostridium perfringens toxin vaccine in China. In addition, while a mixed vaccine is prepared by the vaccine with other antigens, the mixed vaccine can be prepared without increasing a using dose of the mixedvaccine.

The invention discloses a novel coronavirus SARS-CoV-2 replicon as well as a construction method and application thereof, and belongs to the technical field of virology and molecular biology. The novel coronavirus SARS-CoV-2 replicon is an SARS-CoV-2-GFP replicon with a single reporter gene or an SARS-CoV-2-GFP-Luc replicon with double reporter genes. The invention also discloses the constructionmethod and application of the novel coronavirus SARS-CoV-2 replicon. According to the novel coronavirus SARS-CoV-2 replicon disclosed by the invention, infectious virus particles are not generated, sothat the biological safety risk in the process of screening antiviral drugs by utilizing the infectious virus particles is effectively reduced. Besides, a reporter gene carried by the replicon can allow high-throughput screening to be performed, so that the screening efficiency of anti-novel coronavirus drugs is greatly improved.

The invention discloses a closed full-automatic nucleic acid extraction and detection system based on a CRISPR technology. The closed full-automatic nucleic acid extraction and detection system comprises a bag type biological chip and a control detector, wherein the bag type biological chip comprises a nucleic acid extraction cavity, a No.1 cleaning liquid cavity, a No.2 cleaning liquid cavity, aNo.3 cleaning liquid cavity, an eluent cavity, a waste liquid cavity, an amplification cavity and a CRISPR detection cavity which are respectively used for storing various liquids required by nucleicacid extraction and detection, and the cavities are connected through micro-channels; and the control detector comprises a push rod controller, a controllable alternating electromagnet and a fluorescence detector, wherein the push rod controller is used for controlling a flow direction of liquid, the controllable alternating electromagnet is used for stirring and adsorbing magnetic beads, and thefluorescence detector is used for CRISPR luminescence detection of nucleic acid. According to the closed full-automatic nucleic acid extraction and detection system based on the CRISPR technology, nucleic acid cross contamination during detection can be avoided, in addition, the bag type biological chip and the control detector are relatively separated, a detection sample can be rapidly replaced,and practical detection application is facilitated.

The invention relates to a brassica napus ALS (Acetolactate Synthase) gene promoter and application and belongs to the technical field of biology. The brassica napus ALS gene promoter disclosed by the invention is characterized in that a brassica napus ALS gene promoter sequence is colonized; a promoter element contained in the sequence is predicted by bioinformatics software; the activity of the gene promoter is subjected to qualitative detection and analysis by constructing a plant expression vector; and the results show that the colonized sequence can drive a GUS reporter gene to be expressed in all organs of tobacco seedlings, has high expression activity and characteristics of a constitutive promoter and can be applied to plant genetic engineering research as the strong constitutive expression promoter. The promoter can replace a cauliflower mosaic virus (CaMV) 35S promoter and an agrobaeterittm tumefaciens nopaline gene nos promoter which are normally used in the current plant genetic engineering research to construct a plant transgenic expression vector; and the plant transgenic expression vector is applied to genetic engineering of dicotyledons, so that the biological safety risk of transgenic plants is reduced.

The invention discloses a quick time-resolved fluorescence immunoassay kit for detection of T cells infected with tuberculosis. The kit is characterized by comprising 1) a test reaction cup; 2) a calibrator reaction cup; 3) a blank control culture tube containing an anticoagulant substance inside; 4) a test culture tube; 5) a positive control culture tube containing an anticoagulant substance and cell agglutinin inside; 6) an experiment buffer solution and 7) a wash concentrate. The invention further discloses a detection method of the quick time-resolved fluorescence immunoassay kit for detection of T cells infected with tuberculosis. The kit adopts a gamma-interferon in-vitro release method, a biotin-avidin method and double-antibody sandwich time-resolved fluorescence immunoassay together, the experimental methods are simple and quick, automatic operation is realized, and the detection system is an open type operation system.

The invention relates to a polycyclic aromatic hydrocarbon degradation gene engineering strain. The strain Pseudomonas putida GLEB3 is collected by China General Microbiological Culture Collection Center on September 19th, 2016; the collection number is CGMCC No.13014; and the collection address is Institute of Microbiology, Chinese Academy of Sciences, No.1 Yard 3, Beichenxi Road, Chaoyang District, Beijing City. The controllability of the gene engineering strain can be effectively enhanced in the growth process, and the biosafety risk is lowered in the microbe-reinforced restoration process.

The invention relates to the fields of molecular virology and genetic engineering. In particular, the invention relates to an HIV (human immunodeficiency virus) envelope protein, a coding sequence thereof, and an HIV pseudovirus comprising the HIV envelope protein; the HIV pseudovirus can be used to detect ADCC (antibody-dependent cell-mediated cytotoxicity) activity of an anti-HIV antibody. The invention also relates to a kit for detecting the ADCC activity of the anti-HIV antibody. The invention further relates to a method for detecting the ADCC activity of the anti-HIV antibody by using the HIV pseudovirus, the method can achieve rapid, simple and high-throughput detection of the ADCC activity of the anti-HIV antibody, and thereby the method is of great significance for protection effect on analysis of vaccine immune response, and development and quality control of a new vaccine.

The invention relates to a sheep aphthosis virus fusion protein rB2LcF1L. An amino acid sequence of the sheep aphthosis virus B2L and F1L fusion protein is SEQ ID No:2; compared with the wild type sheep aphthous virus F1L protein, the sheep aphthosis virus fusion protein rB2LcF1L has amino acid sequences of sites No.1 to 70 deleted to facilitate the expression of the fusion protein and increase the expression level. At the same time, 6 groups of histidine tags are added to the C-terminus of the fusion protein to facilitate purification in subsequent production. In the above amino acid sequence, compared with a B2L amino acid sequence of a conventional isolate, the V/L of the amino acid at a site No.9 is mutated to G, namely ''IPVG or IPLG is mutated to IPGG'', and the LDCF at a site No.35is mutated to LYCF. The above mutation significantly increases the expression level and immunogenicity of the fusion protein.

The invention provides an automatic nucleic acid extraction detector and a detection method thereof. The automatic nucleic acid extraction detector comprises a chip and a hardware system, wherein the chip comprises a supporting body and a plurality of containing cavities with elastic outer walls, the containing cavities are formed in the supporting body, the containing cavity comprises a sample pool, a cracking pool, a washing solution pool, an LAMP buffer solution pool, an amplification detection pool and a waste liquid pool, and the hardware system comprises a side extrusion device for inserting the chip, a temperature control module for adjusting a temperature of each containing cavity, a magnetic force control element for controlling a movement direction of magnetic beads, a fluorescence detection module for detecting nucleic acid and a microcontroller for regulating and controlling operation of the automatic nucleic acid extraction detector. The nucleic acid detection method comprises steps of chip insertion, lysis, washing, elution, amplification, detection and the like. According to the automatic nucleic acid extraction detector and the detection method, cross contamination during nucleic acid detection is avoided, a detection sample can be rapidly replaced, and automatic detection of nucleic acid is facilitated.

The invention relates to the field of genetic engineering, and particularly relates to a vector for analyzing the expression specificity of a plant promoter, a preparation method and application thereof. A T-DNA region of the vector sequentially comprises a screening marker gene expression cassette driven by a callus specific promoter and a reporter gene expression cassette with multiple cloning sites from the 5' end to the 3' end; and the screening marker gene expression cassette driven by the callus specific promoter comprises the callus specific promoter, a screening marker gene coding sequence and a terminator. The vector utilizes the rice callus specific promoter to drive the expression of a screening marker gene, can greatly reduce the non-specific interaction of a target gene promoter, improves the specificity of target gene expression, can effectively reduce the biological safety risk caused by the screening marker gene in transgenic plants, and has important application value in basic theoretical research and molecular breeding.

The invention provides a porcine circovirus gene modified attenuated strain as well as a construction method and application thereof, and relates to the technical field of biological gene engineering. The genome of the attenuated strain is subjected to codon mutation or codon optimization or gene truncation and the like, so that the gene does not encode or encode part of capsid protein (Cap protein) finally, and a fluorescent protein gene is inserted into a nucleotide sequence deleted by gene truncation. On the basis, a genome of the gene modified attenuated strain is transfected into a donor cell capable of expressing capsid protein, and a complete virion capable of infecting a host cell is finally self-assembled by utilizing a replication system of the cell and the genome of the virus.

The invention discloses a method for preparing circular DNA (Deoxyribose Nucleic Acid) in vitro. The method comprises the following steps: designing a Gibson assembly connection fragment according to a target linear DNA fragment, introducing a recognition site sequence of restriction enzyme Bsa I, mixing the target linear DNA fragment and the Gibson assembly connection fragment, carrying out Gibson assembly, carrying out rolling circle amplification and purification by taking a Gibson assembly product as a template to obtain an RCA product, carrying out Bsa I enzyme digestion on the purified RCA product, and carrying out purification to obtain the RCA. Carrying out agarose gel electrophoresis after enzyme digestion, carrying out gel cutting to recover an enzyme digestion band consistent with the target linear DNA fragment in size, and finally cyclizing and purifying an enzyme digestion product by using T4 DNA ligase to obtain a cyclized product of the target linear DNA fragment. The method can replace traditional plasmids, only an incision enzyme recognition sequence with the length of 11 basic groups is additionally introduced except necessary elements, the molecular weight of circular DNA is reduced to the minimum, meanwhile, the biological safety risk is reduced, and in-vitro cyclization of linear DNA fragments is achieved.

The invention discloses a brassica napus Bna.A08IDD7 gene promoter and application thereof, the nucleotide sequence of the promoter is shown as SEQ ID NO.5, and the promoter can be strongly expressedin roots, leaves, stems, flowers, siliques, seeds and other tissues of arabidopsis thaliana; therefore, it is indicated that the sequence has very strong expression activity and characteristics of a constitutive promoter, and can be used as a strong constitutive promoter to be applied to plant genetic engineering research. Therefore, the promoter creates excellent germplasm resources and other genetic engineering research aspects; the promoter has good application potential, can replace a cauliflower mosaic virus (CaMV) 35S promoter commonly used in current plant genetic engineering research,constructs a plant transgenic expression vector, is applied to plant genetic engineering, and can widely cultivate transgenic plants with high biosafety.

The invention discloses a production early warning method and system for a biological product, and relates to the field of artificial intelligence, in particular to the production early warning method and system for the biological product. The method comprises the steps of obtaining a first feature information set by traversing a first biological product; performing hierarchical clustering analysis to generate a first clustering result, and obtaining a first production area set; collecting first production area set sensing information to obtain a first sensing information collection result, and obtaining a first detection result; and generating a first early warning signal for the abnormal feature information. The problems that in the prior art, when pathogenic microorganism biological products are produced, risks of all production links cannot be found in time, targeted measures cannot be taken in time, and finally production and life safety are affected are solved. Through comprehensive and timely monitoring and analysis of biological safety risks and timely early warning of abnormal conditions, the technical effects of improving the production efficiency of biological products, reducing the biological safety risks and guaranteeing property and life safety are achieved.

The present invention relates to a non-toxic clostridium perfringens beta-toxin genetic engineering vaccine. The prepared clostridium perfringens beta-toxin recombinant subunit vaccine is produced bycodon-optimized production and multiple-amino-acid-mutation- containing recombinant clostridium perfringens beta-toxin proteins, thus maximally retains integrity and spatial conformation of natural toxin proteins, keeps immunogenicity, and also avoids biosafety hazards brought by single amino acid mutations. The vaccine also has advantages of simple preparation technology, low immune dose, excellent vaccine efficacy, etc., greatly reduces biosafety risks in vaccine production processes compared with current commercial clostridium perfringens natural toxin inactivated vaccines in China, and isan ideal candidate vaccine for upgrading of current type B and type C clostridium perfringens toxin vaccines in China; and when the vaccine and other antigens are commonly used to prepare a combined vaccine, dose of the combined vaccine cannot be increased and the combined vaccine can be prepared.

The invention discloses a production method of a recombinant strain expressing ETX-CSA at the same time and a vaccine related to the recombinant strain. A construction method of the recombinant straincomprises the following steps of introducing encoding gene of a fusion protein ETX-CSA into escherichia coli to obtain the recombinant strain expressing the fusion protein, wherein an amino acid sequence of linking peptide is 297th-311st sites of SEQ ID No.1. According to the method provided by the invention, two kinds of toxin proteins can be acquired at the same time by fermenting one strain only, and the strain can prevent two diseases after being prepared into a vaccine; after the strain and a C type clostridium perfringens toxin are prepared into a combined vaccine, the combined vaccinecan generate excellent immune effect, and titers of rabbit serums in three components (S, C, D) can be increased to 3, 4 and 10 times of a norm standard respectively. The method provided by the invention is good in stability, short in consumed time and low in cost, wherein the costs of the S component and the D component are lowered to be 1/100 of the traditional technology, and the titer cost ofthe three components (S, C, D) can be increased to be 344, 4 and 993 times of a traditional technology respectively.