1398 results about "Influenzavirus" patented technology

The invention relates to the technical field of biomedicine, in particular to an anti-influenza A virus NP protein antibody pair and application thereof. The antibody pair comprises two monoclonal antibodies; the monoclonal antibody comprises a heavy chain variable region and a light chain variable region; one monoclonal antibody in the monoclonal antibody pair is named as a 10G10 antibody, and the other monoclonal antibody is named as a 11F8 antibody. The invention also discloses a nucleic acid molecule for coding the monoclonal antibody and a hybridoma cell strain for secreting the monoclonal antibody. In addition, the invention also discloses a kit for detecting influenza virus protein, which comprises the monoclonal antibody pair, the 10G10 antibody is used as a capture antibody, and the 11F8 antibody is used as a labeled antibody. The antibody for resisting the influenza A virus NP protein provided by the invention has relatively good specificity on the influenza virus protein.

The invention discloses monoclonal antibodies NP5 and NP8 for resisting influenza A virus NP protein and application of the monoclonal antibodies NP5 and NP8, and belongs to the technical field of monoclonal antibodies. According to the invention, 19 influenza A virus NP protein monoclonal antibodies are prepared, and the monoclonal antibodies NP5 and NP8 are obtained by screening through ELISA (enzyme-linked immuno sorbent assay) detection and sequencing analysis. The NP5 comprises a light chain CDR2 of which the amino acid sequence is RTS, light chains CDR1 and CDR3 of which the amino acid sequences are shown as SEQ ID NO.3-4, and heavy chains CDR1-3 of which the amino acid sequences are shown as SEQ ID NO.7-9; the NP8 comprises a light chain CDR2 of which the amino acid sequence is LVS, light chains CDR1 and CDR3 of which the amino acid sequences are as shown in SEQ ID NO.12 to SEQ ID NO.13, and heavy chains CDR1-3 of which the amino acid sequences are as shown in SEQ ID NO.16 to SEQ ID NO.18. The invention lays a foundation for preparing an influenza A virus detection kit.

The invention provides a nucleic acid composition for respiratory tract pathogen detection and application thereof, and relates to the technical field of biology. The invention provides a nucleic acid detection composition which has high specificity and can be used for detecting various respiratory pathogens. The composition can be used for detecting influenza B virus, influenza A virus H1N1, influenza A virus H3N2, novel coronavirus N gene, novel coronavirus O gene, respiratory syncytial virus, I-type parainfluenza virus, II-type parainfluenza virus, III-type parainfluenza virus, IV-type parainfluenza virus, rhinovirus, adenovirus, haemophilus influenzae, and the like. The kit has 15 pathogens including streptococcus pneumoniae and staphylococcus aureus, the lower detection limit can be as low as 20 copies / reaction, and the kit plays an important role in rapid detection of respiratory tract pathogens.

The invention provides a method for preparing a chicken immortalized cell line. The method comprises the following steps: constructing a plasmid system of conditional over-expression TERT (Telomerase Reverse Transcriptase), carrying out ovarian injection and breeding to obtain a transgenic chicken of the conditional over-expression TERT, and carrying out separation and induction to obtain the chicken immortalized cell line. The obtained chicken immortalized cell line has the characteristics of primary cells and is susceptible to viruses; the method can be used for virus reproduction and detection, influenza virus vaccine production and the like, can obtain immortalized cell lines of all organs and all parts of the chicken in a short time, and has a good application prospect in the vaccine industry.

Methods for the rapid detection of the presence or absence of SARS-CoV-2, influenza A and influenza B in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers and probes targeting SARS-CoV-2, influenza A, and influenza B and kits are provided that are designed for the detection of SARS-CoV-2, influenza A and influenza B.

The invention discloses a primer group and a kit for simultaneously detecting nucleic acids of various respiratory pathogens based on twenty-six PCR (Polymerase Chain Reaction). The kit comprises primer pool freeze-dried powder of influenza A virus (FluA), influenza B virus (FluB), respiratory syncytial virus (RSV), human parainfluenza virus (hPIV), mycoplasma pneumoniae (MP), novel coronavirus (SARS-CoV-2), human respiratory adenovirus (HADV), human respiratory nose / enterovirus (HRV / HEV), bordetella pertussis (BP), human coronavirus (HCoV), human bocavirus (HBoV) and chlamydia pneumoniae (CP), PCR (polymerase chain reaction) freeze-dried powder and a buffer solution. According to the kit, primer system optimization is carried out on various pathogens, brand new primers and probe pools are designed, clinical sensitivity and specificity can be achieved, and meanwhile, the kit is matched with a fully-closed full-automatic detection card box integrating sample treatment and amplification and is suitable for rapid detection of respiratory tract pathogens.

The invention belongs to the technical field of biology, and particularly relates to a group of paired monoclonal antibodies for resisting H9N2 subtype avian influenza viruses and a surface enhanced Raman spectroscopy rapid detection test strip established based on the antibodies. The paired monoclonal antibody comprises a labeled antibody 1E9 and a capture antibody 3C10 which are prepared by taking hemagglutinin protein of the H9N2 subtype avian influenza virus as antigens and can be specifically combined with the H9N2 subtype avian influenza virus. According to the invention, an SERS (Surface Enhanced Raman Scattering) probe Au-coated Pt-4ATP is used as a marker and is coupled with a labeled antibody 1E9 to prepare a labeled probe, and the labeled probe is paired with a capture antibody 3C10 to establish an SERS immunochromatography test strip. The test strip has a colorimetric and Raman dual interpretation mode, can be used for qualitative and quantitative detection, has the advantages of strong specificity, high sensitivity, good repeatability and stability and the like, is simple and rapid in clinical detection operation, and provides reliable technical support for on-site rapid screening of subtype viruses.

The invention discloses a kit for visually detecting an H5 subtype avian influenza virus based on an RPA-CRISPR / Cas12b system and application of the kit, and belongs to the technical field of molecular detection. The kit comprises an RT-RPA (Reverse Transcription-Recombinase Polymerase Amplification) primer pair and sgRNA1 The RT-RPA primer pair comprises a forward primer with a nucleotide sequence as shown in SEQ ID NO.1 and a reverse primer with a nucleotide sequence as shown in SEQ ID NO.2; and the nucleotide sequence of the sgRNA1 is as shown in SEQ ID NO. 7. According to the kit, nucleic acid amplification and detection are synchronously completed through a tubular reaction system, the detection sensitivity reaches 1 copy / mu L, the coincidence rate with fluorescent RT-PCR is 100%, and no cross reaction exists on H7, H9, NDV and IBV. The kit does not need a complex instrument, is suitable for rapid screening on a basic level site, and provides an efficient solution for prevention and control of avian influenza.

The invention belongs to the technical field of vaccines and biology, and particularly relates to a broad-spectrum influenza A vaccine immunogen composition and application. The invention provides a nucleic acid molecule for coding an immunogenic peptide, wherein the immunogenic peptide comprises: M2 protein extracellular region (M2e) peptide fragments which are connected in series; a hemagglutinin protein stem region extracellular domain (HA2) peptide fragment derived from an H7N9 influenza virus subtype; multimerization of the motif peptide fragment; and optionally, one or more linker peptide fragments independently located between the peptide fragments, wherein the polynucleotide molecule comprises a coding nucleic acid of each peptide fragment. The broad-spectrum influenza A vaccine can induce an organism to generate antigen-specific immune response, a high-titer antibody can be generated 14 days after last inoculation, the broad-spectrum influenza A vaccine can completely protect and resist attack of various subtype influenza A viruses, and the broad-spectrum influenza A vaccine has a wide application prospect.

The invention provides a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas-based kit for isothermal rapid visual detection of H1N1 influenza virus and mycoplasma pneumoniae and a detection method thereof. The kit and the detection method provided by the invention can be used for timely, sensitively and specifically detecting a nucleic acid target in a sample. The invention provides a lateral flow analysis (LFA) detection platform of a three-stage amplification system integrating recombinase polymerase amplification (RPA), CRISPR / Cas12a and catalytic hairpin assembly (CHA) circulation, the detection platform is used for sensitively and accurately detecting mycoplasma pneumoniae DNA (MP DNA) and influenza A virus subtype H1N1RNA (H1N1RNA), and compared with other researches based on an LFA-CRISPR / Cas12a system, the detection platform has the advantages that the detection efficiency is greatly improved, and the detection cost is reduced. According to the kit and the detection method provided by the invention, nucleic acids of two pathogens can be visually detected on a single LFA at the same time, and the sensitivity is high.

The invention belongs to the field of biology, and discloses a monoclonal antibody of avian influenza virus PB2 protein, the monoclonal antibody comprises a heavy chain variable region and a light chain variable region, and can specifically bind to influenza virus matrix protein PB2, and the amino acid sequence of the heavy chain variable region is as shown in SEQ ID NO.1; the amino acid sequence of the light chain variable region is as shown in SEQ ID NO. 2; the hybridoma cell has relatively high sensitivity and specificity, and can be specifically combined with at least five subtype avian influenza PB2 proteins; meanwhile, the invention also provides application and drugs of the labeled compound, the hybridoma cell and the monoclonal antibody.

The present disclosure relates generally to multisubunit nucleic acids comprising a plurality of polynucleotide sequences, wherein each polynucleotide sequence of the plurality comprises a target sequence, a linker sequence, and a self-assembling sequence, or a linker sequence, a target sequence, a linker sequence, and a self-assembling sequence, or a combination thereof, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by a cleavage sequence, and wherein the multisubunit nucleic acid further comprises a signal sequence upstream of one or more of the polynucleotide sequences of the plurality, wherein the target sequence is obtained or derived from a respiratory syncytial virus, a metapneumovirus, a human parainfluenza virus, or a combination thereof. The multisubunit nucleic acid encodes a multisubunit peptide.

The invention provides an influenza A / B virus sandwich method detection reagent. The influenza A / B virus sandwich method detection reagent comprises a pair of influenza A virus antibodies and a pair of influenza B virus antibodies, the pair of influenza A virus antibodies comprises a labeled antibody 19C7 and a coated antibody 27H4, the heavy chain amino acid sequence of the 19C7 antibody is SEQ ID NO: 1, and the light chain amino acid sequence of the 19C7 antibody is SEQ ID NO: 2; the heavy chain amino acid sequence of the 27H4 antibody is SEQ ID NO: 9, and the light chain amino acid sequence of the 27H4 antibody is SEQ ID NO: 10; the pair of influenza B virus antibodies comprises a labeled antibody 10G11 and a coated antibody 23E2, the heavy chain amino acid sequence of the 10G11 antibody is SEQ ID NO: 17, and the light chain amino acid sequence of the 10G11 antibody is SEQ ID NO: 18; the heavy chain amino acid sequence of the 23E2 antibody is SEQ ID NO: 25, and the light chain amino acid sequence of the 23E2 antibody is SEQ ID NO: 26. The sandwich method detection reagent has extremely high specificity and sensitivity.

The invention provides a capture probe for detecting a human parainfluenza virus whole genome and a kit and application thereof, and relates to the technical field of gene sequencing. The capture probe composition disclosed by the invention comprises probes of which the sequences are shown as SEQ ID NO. 1 to SEQ ID NO. 947. The capture probe composition disclosed by the invention can fully cover the full-length genome of the human parainfluenza virus type 1-4, and has the characteristics of uniformity, specificity and high capture efficiency on a captured region. According to the method, the whole genome sequence of the human parainfluenza virus can be more economically and efficiently obtained through probe hybridization capture and next-generation sequencing technologies, and the purposes of human parainfluenza virus variation monitoring and virus type traceability can be achieved depending on sequence information.

The invention relates to the technical field of bird flu host species identification, in particular to a new bird flu high-risk host species identification method based on virus sharing entropy. According to the method, on the basis of spatial distribution prediction of bird flu host species, the bird flu virus sharing entropy of the bird flu host species is calculated species by species according to the overlapping areas of the target host species and all other host species by extracting the distribution area overlapping area of the target host species and any other host species, and the bird flu virus sharing entropy of the bird flu host species is obtained. And quantitatively analyzing the possibility that the target host species and other host species are subjected to avian influenza cross-species transmission infection. By comparing and analyzing the entropy value of the shared entropy of the avian influenza virus, the avian influenza host species with high virus reassortment risk are identified, and a scientific basis is provided for prevention and control of epidemic diseases.

The invention discloses a biologically safe and degradable cerium-based tannic acid nano-enzyme as well as a preparation method and application thereof. The nano-enzyme can be administrated through a non-invasive nasal cavity. In an area with excessive ROS, thioketone on CeTA-k1tkP is destroyed, PEG is removed, hydrogen bonds of an amino acid sequence are allowed to form beta-sheet, the beta-sheet is accumulated at an inflammation part, and a fiber or network structure is formed. Meanwhile, CeTA-k1tkP shows enzyme-like capacity, ROS is effectively eliminated, inflammation is effectively relieved, and systemic immunosuppression is avoided at the same time. In an influenza A (Flu A) virus or Sendai virus (SeV) mediated viral pneumonia model, CeTA-k1tkP can generate responsive deformation in inflammatory lung tissues, so that the nano-enzyme effectively decomposes ROS (reactive oxygen species). In addition, the CeTA-k1tkP can relieve bacterial inflammation and reduce tissue damage in a mouse viral pneumonia model with secondary bacterial infection.

The invention provides application of a porcine parainfluenza virus type 5 as an expression vector, and belongs to the technical field of biological vaccines. According to the invention, a PIV5 strain separated from diarrhea pigs in Jiangsu Province in China is utilized to construct PIV5 virus infectious clone. The eGFP gene is inserted into a PIV5 infectious cloned virus genome, the recombinant PIV5 virus capable of expressing the green fluorescent protein is successfully rescued, and when the eGFP gene is inserted into a P / M gene spacer region, the expression quantity is the highest, which indicates that an exogenous gene expression cassette can influence the expression level of an exogenous gene, and the P / M gene spacer region is the optimal site for inserting the exogenous gene, so that the recombinant PIV5 virus can be successfully rescued. And a solid foundation is laid for developing a vector vaccine by utilizing the swine PIV5. The porcine parainfluenza virus type 5 can be used as an expression vector to express foreign proteins, can also be used for preparing oral vaccines, and can relieve the stress reaction of pigs in the immune process.

The application belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody combination for detecting canine parainfluenza virus NP protein and application thereof. The combination is composed of monoclonal antibodies 5H10 and 1H7, 5H10 is used as a coating antibody, and 1H7 is used for colloidal gold labeling, so that efficient and specific sandwich detection of CPIV / NP antigen can be realized. The variable region CDR sequences of the heavy chains and light chains of the two antibodies are clear (SEQ ID NO. 1-12). The application also provides a rapid detection test strip based on the antibody combination, which can be used for on-site screening, early diagnosis and epidemiological monitoring of canine parainfluenza virus, solves the problem that existing detection technologies lack high-specificity recognition tools, and has a good application prospect.

The invention discloses primers and a kit for rapidly detecting canine respiratory tract pathogens and a preparation method, and belongs to the technical field of biology. The kit comprises five groups of specific primers for rapidly detecting the canine respiratory pathogens, can rapidly and accurately detect the five respiratory pathogens of canine parainfluenza virus, canine distemper virus, canine adenovirus type II, canine crown virus and canine bordetella within 15 minutes, and is low in detection cost and low in detection environment requirement. The kit can be applied to clinical rapid diagnosis and on-site screening to timely diagnose infection, so that veterinarians can rapidly take effective treatment measures; the technical problems of short time consumption, high accuracy, low cost and simple operation cannot be simultaneously considered in the detection process in the prior art are solved.

The invention belongs to the technical field of biological pharmacy, and particularly relates to influenza virus-like particles for chimeric expression of new coronavirus RBD protein as well as a preparation method and application of the influenza virus-like particles. The virus-like particle is prepared by embedding fusion protein on the surface of influenza virus M1, and the fusion protein comprises an HA2 segment of HA protein of influenza virus, an extracellular functional region of M2e matrix protein, a conserved region short peptide of nucleoprotein, and RBD tandem protein with mutated five sites of 417, 452, 478, 484 and 501 of new coronavirus; the virus-like particles can be used for preventing specific influenza and new crowns; the preparation method of the virus-like particles is simple and low in cost, the virus-like particles are good in immunogenicity and cross protectiveness, the preparation requirements of large-scale influenza vaccines can be met, the biological safety is high, and the immunity of an organism to various influenza viruses and new coronal viruses can be stimulated.

The application provides a virus combined detection immunochromatography test paper, a detection pen and a detection product, and relates to the technical field of immunochromatography test paper. The virus combined detection immunochromatography test paper can detect the novel coronavirus, the influenza A virus and the influenza B virus in a three-in-one manner. The sample binding pad of the immunochromatography test paper is coated with labeled antibodies, including a first influenza A virus antibody, a first influenza B virus antibody, a first novel coronavirus antibody and a first human internal reference protein antibody, and each labeled antibody is labeled with a different color of label. The immunochromatography test paper adopts a double-antibody sandwich principle, and the second influenza A virus antibody, the second influenza B virus antibody and the second novel coronavirus antibody are coated on different detection lines in the detection area. The immunochromatography test paper and the detection pen solve the problems of complex operation and difficult interpretation of the three-in-one detection product of the novel coronavirus, the influenza A virus and the influenza B virus in the prior art.

The invention provides a construction method and application of a recombinant herpesvirus turkey for co-expressing an NDV (Newcastle Disease Virus) F gene and an H9 subtype AIV (Avian Influenza Virus) HA gene, and belongs to the field of genetic engineering vaccines. The recombinant clone technology is utilized, an expression framework containing a Newcastle disease virus gene VII type strain F gene and an H9 subtype avian influenza virus HA gene is inserted between 95322-95323 nucleotides or between 112071-112088 nucleotides of a turkey herpesvirus FC126 strain, and the recombinant clone with the F gene and HA gene expression framework inserted into a turkey herpesvirus genome is constructed and obtained. Recombinant turkey herpesvirus vaccine strains H20901 and H20902 for co-expressing the Newcastle disease virus F gene and the H9 subtype avian influenza virus HA gene are rescued and obtained by the recombinant turkey herpesvirus vaccine strain H20901 and H20902. The vaccine prepared from the recombinant turkey herpesvirus provided by the invention can provide dual protection for the Newcastle disease virus and the avian influenza virus, and has a good market application prospect.

The invention discloses a multi-subtype influenza A virus nucleic acid detection kit and a detection method, and relates to the technical field of medical detection.The kit comprises a group of reverse transcription RT-RPA amplification primer pairs used for amplifying influenza A virus M gene target sequences, the reverse transcription RT-RPA amplification primer pairs are designed for specific areas of subtypes H1N1, H2N2, H3N2, H5N1 and H7N9, and the reverse transcription RT-RPA amplification primer pairs are designed for specific areas of subtypes H1N1, H2N2, H3N2, H5N1 and H7N9; the crRNA library comprises a plurality of crRNA molecules which are respectively complementary with different conserved regions in the M gene target sequence and is used for guiding Cas12a protein to carry out specific recognition; a Cas12a protein reagent; the reporter molecule can be subjected to non-specific cleavage by the Cas12a protein; the enzyme preparation, the buffer solution and the nucleotide substrate are required by RT-RPA reaction. The invention aims to realize synchronous, rapid, high-sensitivity and high-specificity detection of various subtype influenza A viruses.

The invention discloses a serum-free full-suspension MDCK (Madin-Darby Canine Kidney) cell and application thereof in preparation of influenza vaccines, and belongs to the field of virology and biological medicines. The serum-free full-suspension MDCK cell is preserved in the China Center for Type Culture Collection, is classified and named as a canine kidney cell MDCK-JHS, and has a preservation number of CCTCC NO: C2024373. The invention further provides a method for culturing the influenza virus by using the cell line, high-titer amplification of the influenza virus can be realized, and the method is suitable for industrial production.

The invention provides oral streptococcus ZRSOR-5 as well as a related product, a method and application thereof. The preservation number of the streptococcus oral cavity ZRSOR-5 provided by the invention in the Guangdong Microbial Culture Collection Center is GDMCC No: 65352, and the preservation number of the streptococcus oral cavity ZRSOR-5 provided by the invention is CGMCC No: 65352. Experiments prove that the SOR preparation is prepared by diluting the SOR preparation with normal saline and inhaling the SOR preparation into mice in advance, so that the survival rate of influenza virus infected mice can be remarkably increased, and lung inflammation and virus load can be reduced. When the ZRSOR-5 preparation provided by the invention is used in advance, pneumonia caused by influenza viruses can be resisted.

The invention provides a bovine parainfluenza virus type 3 QL3 strain, and the preservation number of the bovine parainfluenza virus type 3 QL3 strain is CGMCC No.46469. The invention also provides a bovine parainfluenza inactivated vaccine containing the bovine parainfluenza virus type 3 QL3 strain and purified F protein and HN protein of the QL3 strain. The novel bovine parainfluenza virus type 3 inactivated vaccine is obtained by optimizing the antigen components of the bovine parainfluenza virus type 3 inactivated vaccine, and high-level neutralizing antibodies can be generated after immunizing cattle; the safety of the vaccine is improved through process optimization. Safety and potency test results show that after the inactivated vaccine is used for immunizing cattle, no adverse reaction exists, and high-level neutralizing antibodies are generated. Results show that the immune effect of the bovine parainfluenza virus type 3 inactivated vaccine can be improved.

The invention discloses mRNA, an mRNA-LNP vaccine of the mRNA, a preparation method and application of the mRNA-LNP vaccine. The invention discloses an H5 subtype avian influenza virus vaccine, and aims to provide a vaccine which combines a mosaic sequence with an mRNA vaccine, achieves a good effect in an immune protection experiment of chickens, and can realize cross protection of branched strains of H5 subtype avian influenza virus 2.3. 4.4b and 2.3. 4.4 hours. According to the technical scheme, the nucleotide sequence of the mRNA is as shown in SEQ ID NO. 1; the nucleotide sequence of the mRNA is a Mosaic sequence which is obtained by designing a gene sequence of a 2.3. 4.4b branch strain of the avian influenza H5N6; the mRNA-LNP vaccine comprises mRNA (Messenger Ribonucleic Acid) with a nucleotide sequence as shown in SEQ ID NO. 1; belongs to the technical field of biology.

The invention provides a camellia nitidissima plant endophytic bacterium as well as a fungicide and application thereof in resisting influenza A virus. The golden camellia plant endophytic bacteria are bacillus thuringiensis and are preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.32945. A fermentation product of the strain shows good antiviral activity in both in-vitro and in-vivo experiments, and can effectively inhibit replication of influenza viruses and protect infected mice from being damaged by lethal influenza viruses; the compound also shows the effect of regulating immune response, and the secretion of IFN-gamma is enhanced by down-regulating the secretion of proinflammatory factors such as IL-6 and IL-10, so that the immune resistance of the body is improved.

The invention discloses a D / Yama2019 genetic evolution pedigree influenza delta virus reverse genetic operating system. According to the invention, DNA fragments corresponding to segments of PB2, PB1, P3, HEF, NP, M and NS of a full-length genome of D / JY3002 are amplified by using seven pairs of designed primers, the DNA fragments are seamlessly cloned to a pHW2000 vector and a pCC1-DualPro vector to construct plasmids, the plasmids are co-transfected to a cell line, and the artificially rescued D / Yama2019 genetic evolution pedigree influenza delta virus rD / JY3002 is obtained; the genomic sequence of the rD / JY3002 is completely consistent with the genomic sequence of a naturally separated strain D / JY3002, the infection ability is equivalent, and the rD / JY3002 can be stably subcultured for at least five generations. According to the present invention, the constructed unidirectional expression plasmid pPolI-D / JY3002-PB1-240-GFP-240 is adopted to obtain the artificially rescued recombinant influenza delta virus rD / JY3002-GFP carrying the green fluorescence reporter gene, and after the host cell is infected, the GFP expression quantity is high and stable; the invention establishes an efficient and stable D / Yama2019 genetic evolution pedigree influenza delta virus reverse genetic operating system which can be used for developing an influenza delta virus vector presenting exogenous gene.