65results about How to "Strong conservative" patented technology

The invention provides a primer, a probe and a kit for detecting a novel coronavirus. The specific primer and the probe are designed according to an ORF1ab gene, an E gene, an N gene, an S gene and anM gene of the novel coronavirus 2019-nCoV and are used for identifying and detecting the novel coronavirus. The specific primer / probe and the kit for detecting the 2019-nCoV are high in sensitivity,precision and accuracy, good in specificity and stability, small in template amount required by detection and objective and accurate in detection result. The five-target design can well overcome falsenegative caused by variation in the RNA virus passage process, the positive detection rate can be effectively improved, and missing detection is avoided as much as possible. High clinical applicationvalues are realized.

The invention relates to a primer for amplifying novel coronavirus and an application thereof. The primer comprises at least one oligonucleotide primer capable of recognizing a specific region on a novel coronavirus N gene, and the sequence range of the specific region on the novel coronavirus N gene comprises a region sequence of a novel coronavirus reference strain genome 28778-28971. By adopting the primer, novel coronavirus nucleic acid can be rapidly detected from RNA extracted from samples such as blood, saliva, sputum and air; the method has the advantages of simplicity, convenience, quickness and no need of training of large instruments and professionals, can be used as a primary screening and environment detection method for novel coronavirus infected suspected pneumonia cases, and can also be used as an important reference for clinical diagnosis.

The invention relates to a multiple-gene helicobacter pylori detection system and a kit and application thereof. The helicobacter pylori detection system includes 21 pairs of primers respectively for a strain identification gene (16S rRNA), virulence genes (cagA, vacA-s1, vacA-s2, vacA-m1, vacA-m2, iceA1, iceA2, dupA, oipA and luxS), drug resistance genes (2143 locus of 23S rRNA, 148 locus of rdxA, 1777 locus of pbp1A and polymorphism of the 261 locus of gyrA) and quantitative analysis genes ureC and beta-globin of the helicobacter pylori. The multiple-gene helicobacter pylori detection system and the kit of the system do not need the steps including conventional isolated culture and the like, synchronous detection and analysis on strain identification, quantification, virulence and medicine resistance can be directly conducted on tissue samples in the same reaction system, the shortcomings of low flux, long consumed time, low detection rate and the like of a conventional detection method are overcome, a comprehensive, accurate and low-cost etiological diagnosis is clinically provided for the first time, and important references are provided for individualized diagnosis and accurate treatment of helicobacter pylori infection.

The invention relates to a novel guide RNA (Ribonucleic Acid) expression cassette used in a CRISPR / Cas (Clustered Regularly Interspaced Short Palindromic Repeats) system. According to the guide RNA expression cassette, a 5S rRNA gene is adopted as a starter to start expression of guide RNA. The invention further provides a CRISPR / Cas system with the guide RNA expression cassette and a method for editing genomes by using the system. The guide RNA expression cassette, the CRISPR / Cas system and the genome edition method provided by the invention have the advantages of universality, high efficiency, convenience, accuracy and the like.

The invention relates to a system for detecting multiple quantitative and virulent genes of H.pylori as well as kits and applications of the system. The system for detecting multiple quantitative and virulent genes of H.pylori comprises multiple pairs of primers, each for strain identification genes (16S rRNA), quantitative analysis genes ureCand Beta-globin, as well as virulent genes (cagA, vacA-s1, vacA-s2, vacA-m1, vacA-m2, iceA1, iceA2, dupA, oipA and luxS). The system for detecting multiple quantitative and virulent genes of H.pylori and the kits of the system can directly perform synchronous detection and analysis on the strain identification, quantification and virulence of a tissue sample in a same reaction system without adopting a conventional culture step or other steps, remedy defects that a conventional detection method is low in throughput, time-consuming and low in detection rate, obviously improve the accuracy of detection results, immediately provide a comprehensive, accurate and low-cost etiological diagnosis for clinic, and provide an important reference for the accurate diagnosis, differential diagnosis and disease prognosis of H.pylori infection.

The invention relates to construction and screening as well as applications for a siRNA expression carrier of a stomach cancer target STAT3 gene. Two expression plasmids STAT3-siRNA1 / 2 in mammal cells for expressing siRNA are constructed by mainly utilizing an RNA interference technology aiming at different target sequences of an STAT3 gene. The siRNAs expression carrier of the stomach cancer target STAT3 gene provided by the invention can inhibit STAT3 gene expression efficiently with specificity, and can be used for preparing gene medicaments for treating high-expression STAT3 gene stomach cancer.

The present invention discloses an alternating current / direct current microgrid economical dispatching method based on multi-interval nondeterminacy and robust optimization. The method comprises the following steps of: the step 10) obtaining nondeterminacy prediction parameters, and distributing a nondeterminacy time period budget number; the step 20) based on the step 10), constructing a multi-interval nondeterminacy set; the step 30) obtaining an operation cost coefficient and an operation limiting value of each device in the microgrid, and based on the step 20), establishing a robust optimization economical dispatching model of an alternating current / direct current microgrid; and the step 40) solving a robust economical dispatching problem: employing a column constraint generating algorithm to perform iteration solution of the optimization problem, and obtaining the robust operation plan of the alternating current / direct current microgrid. The alternating current / direct current microgrid economical dispatching method can improve the economic benefit of microgrid operation and can improve the important reference for formulation of an operation plan of the alternating current / direct current microgrid.

The invention aims to provide a kit and method for detecting HER2 gene amplification based on a digital droplet PCR technology so as to solve the technical problem of low accuracy of an HER2 gene copynumber variation judgment method in the prior art.

The invention discloses an enteric cytopathogenic human orphan virus 6-type VP1 protein specific epitope, a fusion protein of the enteric cytopathogenic human orphan virus 6-type VP1 protein specific epitope and a preparation method and use of the fusion protein and relates to the field of a gene engineering technology, a vaccine and a diagnostic reagent. The preparation method comprises analyzing an enteric cytopathogenic human orphan virus 6-type surface protein VP1 amino acid sequence by a computer, and screening strong specific epitope-containing protein fragments comprising 75th-101th amino acids, 127th-141th amino acids, 204th-237th amino acids and 257th-289th amino acids, wherein the four protein fragments are connected by 2 glycines and 1 serine so that the epitope fusion protein is formed. A codon in favor with eucaryotes and prokaryotes is used, a brand new gene sequence of the epitope fusion protein is chemically synthesized, through a gene engineering technology, the enteric cytopathogenic human orphan virus 6-type VP1 protein epitope fusion protein is expressed and prepared and is used for development of an enteric cytopathogenic human orphan virus antibody detection reagent and preparation of an enteric cytopathogenic human orphan virus monoclonal antibody and polyclonal antibody.

The invention discloses 14 specific molecular markers for the 1R-7R chromosomes of rye based on a rye EST (expressed sequence tag) sequence, and the molecular markers are forward primers and reverse primers which are designed according to the EST sequence of rye chromosomes; and the molecular markers are used for identifying the specific markers of the 1R-7R chromosomes of rye. The identification of the 1R-7R chromosomes of rye by applying the molecular markers comprises the basic steps of: A. selecting and preparing the marker primers; B. diluting the marker primers; and C. marking the specificity of the rye chromosomes: C-1, extracting template DNA; C-2, carrying out PCR (polymerase chain reaction) amplification; C-3, detecting an amplified product; and C-4, comparing results. In the invention, a set of the specific molecular markers for the rye chromosomes are designed and developed on the basis of the EST sequence of the rye chromosomes, and the whole 1R-7R rye chromosomes in a genetic background of wheat can be rapidly and accurately identified by using a common PCR technology by means of the molecular markers, and thus the invention provides a simple and effective molecular identification method for the transfer of beneficial genes on the rye chromosomes to wheat cultivation.

The invention provides an RT-qPCR detection method, primers and TaqMan probe for detecting a swine transmissible gastroenteritis virus N gene. Particularly, the invention provides a set of RT-qPCR detection method for detecting the swine transmissible gastroenteritis virus gene and a forward primer shown in SEQ ID No.1 in a sequence table, a reverse primer shown in SEQ ID No.2 in the sequence table, and a TaqMan probe sequence shown in SEQ ID No.3 in the sequence table. The invention provides an RT-qPCR detection method for detecting the swine transmissible gastroenteritis virus N gene on the basis of the primers and the TaqMan probe. The detection method has excellent detection sensitivity, and in the case of 1*10<5>copies / microliter to 1*10<1>copies / microliter, Ct values and fluorescence values show typical amplification curves. At the same time, the detection method has good repeatability, strong specificity and convenience in operation.

The invention provides a method for obtaining species-specific consensus sequences of microorganisms, which at least comprises the following steps: S100, searching for candidate consensus sequences: clustering specific sequences of target strains belonging to the same strain based on a clustering algorithm to obtain a plurality of candidate species-specific consensus sequences; S200, verifying andobtaining a species-specific consensus sequence of primary screening; judging whether the candidate species-specific consensus sequences meet the following conditions: 1) the plant seed coverage degree meets a preset value; 2) the effective copy number meets a preset value; if the candidate species-specific consensus sequences meet all the conditions, determining that the candidate species-specific consensus sequences are species-specific consensus sequences. The method is high in sensitivity; a repetitive sequence can be found in an incompletely assembled motif; obtained species-specific consensus sequences are accurate, and the subspecies level can be identified; the conservative property of identified consensus sequences is high, and the maximum value of the plant seed coverage is achieved as much as possible with the least consensus sequences; and all logic modules have multiple verifications, so that the accuracy is high.

The present invention relates to a fast synchronous inspection method of micro content bacteria, which comprises: 1) mould board preparation, 2) PCR augmentation, the augmentation guide object is the general guide object of bacteria ribosomal ribonucleic acid gene, 3) gel electrophoresis of denaturization gradient, i.e. gel electrophoresis of denaturization gradient for augmentation outgrowth to determine sorts. The augmentation of 16S rRNA gene segment of bacteria is realized through UPPCR technology, obtaining all corresponding bacteria gene segment sequence information, and then DGGE is employed to appraise the augmentation outgrowth of mixed bacteria, so as to confirm the inspected bacteria. The invention provides a fast synchronous inspection method of micro content bacteria by combining the technologies of UPPCY and DGGE, which achieves the advantages of micro content, fast and synchronous inspection for mixed bacteria, especially pathogenic bacteria.

The invention discloses an ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for bovine coronavirus. The ELISA detection kit is characterized in that purified BCoV-CD isolate recombinant pET-32a-N protein is used as coating antigen, reaction conditions are optimized, and an indirect ELISA diagnosis method for detecting bovine-serum characteristic N protein antibodies. The ELISA detection kit disclosed by the invention has the beneficial effects that escherichia coli is adopted for prokaryotic expression, the raw-material sources are wide, the preparation is easy, the standardization iseasy, and the production and detection processes are safer and more reliable, so that the ELISA detection kit is suitable for being promoted and used in basic units.

The invention relates to a helicobacter pylori identification and virulence multiplex gene detection system, a kit adopting the detection system and the application of the detection system. The detection system comprises a plurality of pairs of primers aiming at strain identification genes (16S rRNA) and virulence genes (cagA, vacA-s1, vacA-s2, vacA-m1, vacA-m2, iceA1, iceA2, dupA, oipA and luxS) respectively. According to the helicobacter pylori identification and virulence multiplex gene detection system and the kit adopting the detection system, routine culture and other steps are not needed, helicobacter pylori identification and multiplex virulence analysis can be directly conducted on a tissue sample in the same reaction system, the defects of conventional detection methods of low flux, high time consumption and low detection rate are overcome, comprehensive, accurate and low-cost etiology basis is provided for clinical application quickly, and important reference is provided for helicobacter pylori infection accurate diagnosis and differential diagnosis and disease prognosis.

The invention discloses a method for extracting superoxide dismutase from the blood of a mammal, belonging to the technical field of food processing. The method is used to solve the problems of high cost, low benefit and low purity in extracting superoxide dismutase from the blood of the mammal. The method for extracting superoxide dismutase from the blood of the mammal comprises the following steps of: (a) collection of the blood of the mammal; (b) hemolysis; (c) thermal denaturation in the presence of copper salt; (d)ultrafiltration; (e) precipitation of acetone and recovery of acetone with a rotary evaporator; (f)column chromatography; and (g)freeze-drying. The method for extracting superoxide dismutase from the blood of the mammal can be widely used in extracting superoxide dismutase from the blood of any mammal.

The invention discloses a labeled peptide of a sheep-derived component and application of the labeled peptide to detection of gelatin and gelatin products, and belongs to the technical field of detection. A labeled peptide, namely Gly-Pro-Ala-Gly-Pro-Thr-Gly-Pro-Ala-Gly-Lys of the sheep-derived component is involved, and whether the sheep-derived component is contained or not can be determined bydetecting the labeled peptide, specifically, pancreatin digestion is carried out on protein components in a sample to dissociate the labeled peptide, and then whether the labeled peptide is containedor not is detected through a liquid chromatograph-mass spectrometer to judge whether the sheep-derived component is contained or not. A method is strong in characteristic, high in sensitivity and easyto operate, and can determine the sheep-derived component in the gelatin and gelatin products.

The invention discloses a secale cereale EST (expressed sequence tag) sequence based specific molecular marker of a secale cereale 2R chromosome, which comprises a forward primer and a reverse primer designed according to an EST on a secale cereale chromosome, and is the specific marker used for identifying the secale cereale 2R chromosome. The basic steps used for identifying the secale cereale 2R chromosome through the application of the molecular marker are as follows: A, selecting and preparing marked primers; B, diluting the marked primers; and C, the specific marker of the secale cereale 2R chromosome: C-1, extracting DNA of a template, C-2, performing PCR (polymerase chain reaction) amplification, C-3, detecting amplified products, and C-4, performing results comparison. By virtue of the molecular marker provided by the invention, the ordinary PCR technology can be adopted to quickly and accurately identify the secale cereale 2R chromosome in a wheat genetic background, so that a simple, convenient and effective molecular identification method is provided for transferring useful genes on the secale cereale chromosome into a wheat cultivation background.

The invention discloses preparation and application of an echovirus type 1 VP1 protein specific antigen epitope and a fusion protein thereof, and relates to the fields of gene engineering technology, vaccines and diagnostic reagents. An amino acid sequence of the echovirus type 1 surface protein VP1 is analyzed by a computer, and protein fragments, namely a protein fragment from a 75th amino acid to a 140th amino acid and a protein fragment from a 203rd amino acid to a 236th amino acid, containing strong specific antigen epitopes are screened; the two protein fragments are connected by two glycines and one serine, and the antigen epitope fusion protein is formed. Codons preferred by eukaryotes and prokaryotes are selected, and the bran-new gene sequence of the antigen epitope fusion protein is chemically synthesized; with use of the genetic engineering technology, the echovirus type 1 VP1 protein antigen epitope fusion protein is expressed and prepared, is used for development of echovirus antibody detection reagents, and is used for preparation of echovirus monoclonal and polyclonal antibodies.

The invention relates to a coding sequence of green cucumber collateral inhibition gene CLS in genetic engineering technology field. The coding sequence comprises coding polypeptides nucleotide sequence with cucumber collateral inhibition albumen activity, and the nucleotide sequence and 491-1666 bite nucleotide sequence in SEQ ID NO.3 have at least 701mology. The opening reading frame of coding sequence is 1175 bases from 491th to 1666th. The invention can stimulate forming of lateral bud anlage and is used for alimentary crops, floral plants, economic crop and oil crop to increase the number of branch stem thereby raise productivity.

The invention relates to a molecular detection method of salmonella. According to the molecular detection method, a single primer isothermal amplification method is adopted, a specific sequence of a salmonella invA gene is taken as a target sequence, SYBER Green II is taken as a fluorescent dye, and amplification signals are detected through real-time fluorescent quantitative PCR (Polymerase Chain Reaction). The invention further relates to application of the molecular detection method. The detection method disclosed by the invention is high in sensitivity, strong in specificity and sensibility and short in consumed time, has relatively low requirements on template DNA, and is simple and convenient.

The invention discloses poultry-derived characteristic collagen peptide and an application thereof in detection of collagen hydrolysate and products thereof, and belongs to the field of detection. Theamino acid sequence of the characteristic collagen peptide is Gly-Leu-Val-Gly-Gly-Gly-Hyp-Gly-Pro-Ala-Gly-Ala-Lys, and the amino acid sequence of the characteristic collagen peptide is as shown in the specification: Gly-Gly-Gly-Pro-Ala-Gly-Pro-Ala-Gly-Pro-Ala-Gly-Pro-Ala-Gly-Ala-Arg, Val-Gly-Pro-Ile-Gly-Pro-Ala-Gly-Asn-Arg, and Val-Gly-Gly-Gly-Gly-Asn-Arg. A poultry-derived component is containedor not is determined by detecting one or more characteristic collagen peptides. Pancreatic enzyme digestion is carried out on protein components in a sample, so the characteristic collagen peptide isdissociated and is detected by a liquid chromatograph-mass spectrometer. The method has the advantages of strong characteristics and high sensitivity, and can be used for determining the poultry-derived components in the collagen hydrolysate and products thereof.

The invention provides echovirus type-9 VP1 protein specific antigen epitope and a preparation method and application of a fusion protein thereof, and relates to the fields of genetic engineering technology, vaccines and diagnostic reagents. Computer analysis is carried out on the amino acid sequence of the echovirus type-9 surface protein VP1 to screen out protein fragments containing strong specific antigen epitope, namely 75th amino acid-102nd amino acid, 127th amino acid-140th amino acid, 204th amino acid-237th amino acid and 257th amino acid-300th amino acid, and the four protein fragments are connected through two glycines and one serine to form one antigen epitope fusion protein. Codons preferred by both eukaryon and prokaryote are selected for chemical synthesis of a brand new gene sequence of the antigen epitope fusion protein, the genetic engineering technology is utilized to prepare the antigen epitope fusion protein of the echovirus type-9 VP1 protein by expression, and the antigen epitope fusion protein is used for developing an echovirus antibody detection kit and preparing echovirus monoclonal antibody and polyclonal antibody.

The invention discloses a robust predictive control method for a stirring reaction tank, which comprises the following steps: establishing a quasi-linear structure model with a time-varying coefficient; constructing a polyhedron model capable of wrapping the future dynamic state of the stirring reaction tank system by utilizing the boundary information of the time-varying coefficient of the quasi-linear structure model, namely a state space model; and designing a robust prediction controller based on the state space model, and obtaining the optimal control input quantity w1(t) acting on the stirring reaction tank system by using the robust prediction controller, thereby adjusting the input flow w1(t) of a reactant A in real time, and achieving the purpose of controlling the concentration Cb(t) of a product C to track a set value. The robust predictive control method for the stirring reaction tank significantly increases the degree of freedom of the robust predictive controller in the process of optimizing the control rate online.

The invention discloses a chicken-derived characteristic III type collagen peptide and application thereof in detection of collagen hydrolysate and products thereof, which belong to the technical field of detection. Characteristic III type collagen peptide amino acid sequences of the chicken-derived components are Gly-Ala-Gln-Gly-Pro-Pro-Gly-Pro-Thr-Gly-Ala-Arg and Gly-Ser-Hyp-Gly-Pro-Hyp-Gly-Pro-Ser- Gly-Pro-Ala-Gly-Asp-Arg. By detecting one or two peptides, whether the chicken-derived component is contained or not is determined. Pancreatic enzyme digestion on protein components in a sample is carried out to dissociate characteristic III type collagen peptide, whether the characteristic III type collagen peptide is contained or not is detected by using a liquid chromatograph-mass spectrometer, and whether the chicken-derived components are contained or not is judged. The method has the advantages of strong characteristics, high sensitivity and simple operation, and can be used for determining the chicken-derived components in the collagen hydrolysate and products thereof.

The invention discloses a method for inhibiting geminivirus infection by overexpression of arabidopsis ABI5 protein, which inhibits transcription of geminivirus coding genes by overexpression of plant transcription factor ABI5 protein and combination of ABI5 protein and a promoter of geminivirus through specificity, and realizes improvement of virus resistance of plants.

The invention provides a molecular beacon probe and a kit for detecting equine influenza virus pathogens. The molecular beacon probe and the kit belong to the field of biotechnology. The kit comprisesa primer pair for detecting a PB1-D sequence of H3N8 and a molecular beacon probe, nucleotide sequence of upstream primer of primer pair is SEQ: 1; wherein the nucleotide sequence of the forward primer is SEQ: 1, the nucleotide sequence of the reverse primer is SEQ: 2, the nucleotide sequence of the molecular beacon probe is SEQ: 3, the 5'end of the molecular beacon probe is subjected to fluorescence labeling with 5 (6)-carboxyl fluorescein (FAM), and the 3 'end of the molecular beacon probe is subjected to fluorescence labeling with quenched fluorescence 4-(4'-oxane aminobenzene azo) benzoicacid (DABCYL). The kit can inhibit generation of fluorescence signals caused by non-specific binding of the molecular beacon probe and reduce the signal-to-background ratio, so that detection signalscan accurately reflect the number of templates, and the specificity, sensitivity, repeatability and accuracy of detection are improved.

The invention relates to construction and screening of siRNAs (small interfering RANs) expression vectors of a targeted DEC1 (differentiated embryo-chondrocyte expressed gene 1) resulting in gastric cancer and application thereof. In the invention, by mainly adopting an RNA interference technology, two expression plasmids DEC1-siRNA1 / 2 capable of expressing siRNA in mammalian cells are constructed by aiming at different target sequences of the DEC1. The siRNAs expression vectors of the targeted DEC1 resulting in gastric cancer can efficiently and specifically inhibit DEC1 expression; and the siRNAs expression vectors are used for preparing a gene drug for treating gastric cancer resulting from the high expressed DEC1.

The invention relates to a system for detecting multiple drug-resistant and quantitative genes of H.pylori as well as kits and applications of the system. The system for detecting multiple drug-resistant and quantitative genes of H.pylori comprises multiple pairs of primers, eachforstrain identification genes (16S rRNA), drug-resistant genes (polymorphism of a 2143rd locus of 23S rRNA, a 148th locus of rdxA, a 1777th locus of pbp1A and a 261st locus ofgyrA), and quantitative analysis genes ureC and beta-globin. The system for detecting multiple drug-resistant and quantitative genes and the kits of the system can directly perform synchronous detection and analysis on the strain identification, drug resistance and quantification of a tissue sample in a same reaction system without adopting a conventional culture step or other steps, remedy defects that a conventional detection method is low in throughput, time-consuming and low in detection rate, obviously improve the accuracy of detection results, immediately provide a comprehensive, accurate and low-cost etiological diagnosis for clinic, and guide the accurate diagnosis and individualized medication of H.pylori infection and the observation of curative effects.

The present invention relates to a fast synchronous inspection method of micro content bacteria, which comprises: 1) mould board preparation, 2) PCR augmentation, the augmentation guide object is the general guide object of bacteria ribosomal ribonucleic acid gene, 3) gel electrophoresis of denaturization gradient, i.e. gel electrophoresis of denaturization gradient for augmentation outgrowth to determine sorts. The augmentation of 16S rRNA gene segment of bacteria is realized through UPPCR technology, obtaining all corresponding bacteria gene segment sequence information, and then DGGE is employed to appraise the augmentation outgrowth of mixed bacteria, so as to confirm the inspected bacteria. The invention provides a fast synchronous inspection method of micro content bacteria by combining the technologies of UPPCY and DGGE, which achieves the advantages of micro content, fast and synchronous inspection for mixed bacteria, especially pathogenic bacteria.