1113 results about "Influenzavirus" patented technology

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 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 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 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 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 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 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 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.

The invention discloses a preparation method of influenza D virus-like particles, and belongs to the field of veterinary biological products. Influenza D virus (IDV) is one of important pathogens of bovine respiratory disease syndrome (BRDC), the disease causes a hundreds of billions of dollars economic loss to the global cattle raising industry every year, however, commercialized vaccines for influenza D do not exist in the market at present. The preparation method comprises the following steps: target gene amplification, construction of recombinant shuttle plasmids, construction of recombinant baculoviruses, rescue of recombinant baculoviruses, and preparation and purification of virus-like particles. The D-type influenza virus-like particles (IDV VLPs) are obtained through the steps. After mice are immunized, the IDV VLPs are found to have high immunogenicity and can be used as a candidate vaccine for preventing the D-type influenza.

The invention belongs to the field of biological medicine industry, and discloses a blood marker for assisting in diagnosing influenza. The inventor verifies that the LRG1 is a peripheral blood marker of influenza for the first time and can be used for auxiliary diagnosis of influenza and increase of the detection rate of influenza virus by being matched with multiple times of detection of influenza virus. A test result shows that the peripheral blood LRG1 content 5ng / ml is a CUTOFF value, 46 patients and 4 false negative patients are successfully detected for 50 influenza patients, and the sensitivity is 92%; the number of normal persons is 60, the number of the normal persons is 12, and the specificity is 80%. After influenza treatment, LRG1 and Plt are obviously reduced compared with those before influenza treatment; 0.001, which is equivalent to the level of normal people, Pgt; and 0.05%.

The present application provides chimeric proteins comprising an antibody moiety that specifically binds to a component of an influenza virus or a variant thereof, and a positively charged mucoadhesive peptide fragment. Compositions comprising the chimeric proteins described herein are useful for preventing or treating an infection caused by an influenza virus or a variant thereof in an individual.

The present invention uses ex vivo human airway cultures to assess the human transmissibility and replication competence of influenza and coronavirus strains. By comparing pandemic influenza A subtype H1N1 and highly pathogenic avian influenza H5N1 as reference strains, the transmissibility risk of various viruses was evaluated and categorized. Additionally, an in vitro model evaluated virus-induced impairment of alveolar fluid clearance (AFC) as an indicator of disease severity. The study revealed correlations between bronchus viral replication, human transmission, AFC impairment, and clinical disease severity across different influenza and coronavirus strains.

Provided are an anti-neuraminidase human monoclonal antibody obtained by means of multiple rounds of in-vitro screening and recombinant modification, and the use thereof in the preparation of a product for preventing, detecting, or treating influenza viruses. The provided monoclonal antibody exhibits significant neuraminidase inhibitory activity and broad-spectrum binding capacity to influenza virus antigens, shows excellent preventive and therapeutic effects against multiple seasonal influenza viruses, provides a new technical option for resisting influenza infection, and has great application value.

The invention discloses an H9N2 subtype avian influenza virus, the H9N2 subtype avian influenza virus is A / chick / Guangxi / C1228 / 2015 (H9N2), and the preservation number of the H9N2 subtype avian influenza virus is CGMCC (China General Microbiological Culture Collection Center) No.45222; or the H9N2 subtype avian influenza virus is A / quil / Guangxi / 210Q33 / 2015 (H9N2), and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No. 46091. The bivalent inactivated vaccine of CK / GX / C1228 / 15 with better antigenicity and the strain QL / GX / 210Q33 / 15 which has better antigenicity and is separated from quails in the live poultry market has a better protection effect on two different branches of H9N2 subtype avian influenza viruses, the lowest immune dose (0.3 mL) can reach the protection rate of 80% or above, and the generated effective protection antibody can last to 24W.

The invention discloses macromolecular antibacterial artificial leather and a preparation method thereof, and relates to the technical field of artificial leather preparation. The preparation method comprises the following steps: preparing zinc-loaded silicon dioxide with amino grafted on the surface and quaternized polyether polyurethane containing an epoxy group, mixing the zinc-loaded silicon dioxide and the quaternized polyether polyurethane to prepare antibacterial protection slurry, and bonding the zinc-loaded silicon dioxide and the quaternized polyether polyurethane through ring-opening reaction of the amino and the epoxy group during curing to form a cross-linked network, so as to obtain the antibacterial protection layer with antibacterial property, flexibility and wear resistance; zinc ions have an obvious inhibiting effect on bacteria such as gram-positive bacteria and gram-negative bacteria and fungi such as candida albicans, quaternary ammonium salt has an obvious inhibiting effect on influenza viruses, and the zinc ions and the quaternary ammonium salt have a synergistic effect, so that the antibacterial range of the artificial leather is wider, the inhibiting force on germs is stronger, and the antibacterial property of the artificial leather is further improved.

The present invention provides engineered soluble F proteins of paramyxoviruses, such as respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and human parainfluenza virus (hPIV). These engineered proteins are stabilized by specific modifications in the wild-type soluble F sequence, such as substitutions in the 023 chain and / or the introduction of engineered disulfide bonds in the 0 hairpin in the F1 subunit. The present invention also provides a nanoparticle vaccine comprising an engineered soluble F immunogen displayed on a self-assembled nanoparticle. The invention also provides methods of using such vaccine compositions in a variety of therapeutic applications, e.g., methods for preventing or treating viral infections, such as RSV, MPV, and PIV infections.

The invention provides an anti-influenza A virus nano antibody and application thereof, the nano antibody comprises a heavy chain variable region, the heavy chain variable region comprises CDR1, CDR2 and CDR3, and the CDR3 of the heavy chain variable region is selected from an amino acid sequence as shown in SEQ ID NO: 8 or SEQ ID NO: 16. The anti-influenza A virus nano antibody can be combined with various influenza A viruses, including subtype influenza viruses such as H1N1, H2N2, H3N2 and H7N9, has strong broad spectrum, has different binding epitopes of the two nano antibodies, and can be applied to double antibody sandwich method detection, enzyme-linked immunosorbent assay and colloidal gold detection. According to the invention, multivalent transformation is also carried out on the nano antibody to obtain a dodecavalent hexamer cyclic structure nano antibody, so that the application scene of the anti-influenza A virus nano antibody can be further extended.