165 results about "Hypotype" patented technology

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 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 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 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.

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 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.

The invention discloses a primer probe combination and a kit for on-site detection of H5, H7 and H9 subtype avian influenza viruses, and relates to the technical field of gene detection. The invention provides a primer probe combination for detecting H5, H7 and H9 subtype avian influenza viruses on site. The primer probe combination comprises a primer pair and a probe for detecting the H5 subtype avian influenza viruses, the primer pair and the probe are used for detecting the H7 subtype avian influenza virus; the invention discloses a primer pair and a probe for detecting an H9 subtype avian influenza virus. The method for detecting the H5, H7 and H9 subtype avian influenza viruses by adopting the primer probe combination and the kit disclosed by the invention integrates sample pretreatment, nucleic acid extraction, purification, amplification and detection, has better sensitivity and accuracy, is simple, convenient, rapid and time-saving to operate, and plays an important role in on-site rapid detection of research and safety management of the H5, H7 and H9 subtype avian influenza viruses.

The invention relates to the technical field of biology, and discloses sgRNA for knocking out a TNIP2 gene, the nucleotide sequence is shown as SEQ ID NO: 1, blocking of the sgRNA to TNIP2 expression is verified through sanger sequencing and Western Blotting experiments, a TNIP2 knockout cell line and a control cell have no obvious morphological and growth difference, and the sgRNA can be used for knockout of the TNIP2 gene. The sgRNA can inhibit the replication ability of the H1 subtype influenza virus by regulating and controlling the expression of a host gene TNIP2, in addition, the invention further discloses application of the sgRNA, a cell strain with the TNIP2 gene knocked out and a construction method of the cell strain.

This application relates to the field of virus detection technology, specifically disclosing a real-time fluorescent PCR detection kit and method for H10N7 avian influenza virus. The kit includes sample lysis extract, sample washing solution I, sample washing solution II, and reaction premix for detecting H10N7 subtype avian influenza virus; the detection method is an integrated detection method. This application achieves simultaneous detection of H10 and N7 subtype AIVs by designing specific primers and probes. The kit prepared using these primers and probes not only improves detection efficiency but also significantly enhances sensitivity and specificity. The combined use of MAOPA technology and real-time quantitative PCR technology further improves the accuracy of the detection results, making this kit of this application of significant application value in avian influenza prevention and control.

The invention discloses RT-qPCR-HRM (real-time quantitative polymerase chain reaction-high-resolution melting) detection primers and a detection method for identifying five different subtype avian influenza viruses, and belongs to the technical field of biology. The invention discloses an RT-qPCR-HRM detection method for identifying five different subtypes of avian influenza viruses, which comprises the following steps: designing a specific primer based on a highly conserved region of an AIV M gene, and simultaneously carrying out genetic typing on five subtypes of AIV strains H3, H5, H6, H7 and H9 in combination with an HRM technology; by comparing melting curve characteristics of M gene segments of different subtypes of AIV, an AIV detection method with good sensitivity and specificity is established. According to the method, the five AIVs of H3, H5, H6, H7 and H9 can be accurately typed, the defect of missing detection of a traditional AIV detection method is overcome, and the method has a wide application prospect.

The invention belongs to the field of biology, and relates to a broad-spectrum monoclonal antibody aiming at avian influenza virus M1 protein and application of the broad-spectrum monoclonal antibody, the monoclonal antibody has specific reaction with A549 cells infected by avian influenza, has no specific reaction with Newcastle disease virus, duck tembusu virus, goose astrovirus and infectious laryngitis virus, has good specificity, and can be used for preparing the broad-spectrum monoclonal antibody. The 5G9 monoclonal antibody has good reactivity with A549 cells infected by H1-H11 subtype AIV, has good broad spectrum, can be used for detecting M1 protein sub-localization after the AIV infected cells are detected by using the 5G9 monoclonal antibody, and can be used for indicating the infection process of the AIV.

The invention relates to construction and application of H7 subtype and H5 and H7 tetravalent avian influenza DNA vaccines. The invention belongs to the field of molecular biology, and aims to provide a DNA (deoxyribonucleic acid) vaccine for providing efficient immune protection for H7 subtype avian influenza virus. Specifically, the invention discloses an H7 subtype avian influenza virus hemagglutinin protein, which has an amino acid sequence as shown in SEQ ID NO: 5 and / or SEQ ID NO: 6. The invention also discloses polynucleotide for coding the protein, and the polynucleotide has a sequence as shown in SEQ ID NO: 1 and / or SEQ ID NO: 2. The polynucleotide can be used for preparing a vaccine aiming at the current epidemic H7 subtype avian influenza virus. The H5 + H7 tetravalent DNA vaccine is obtained by combining the vaccine with a vaccine aiming at the H5 subtype, and the H5 + H7 tetravalent DNA vaccine can be used for prevention and control of current H5 subtype and H7 subtype avian influenza.

The present invention discloses a detection system for simultaneously detecting avian gyrovirus type 1 and human gyrovirus type 1, and its application, relating to the technical field of biological detection. The detection system includes an RAA amplification primer pair for GyVg1, an RAA amplification primer pair for GyH1, 12-crRNA-3, 13-crRNA-2, an ssDNA probe, an ssRNA probe, LbCas12a nuclease, LbuCas13a nuclease and T7 transcriptase. Based on the conserved genes of GyVg1 and GyH1, the present invention respectively designs specific crRNAs and RAA primers, and by combining the RAA amplification technology with the CRISPR-Cas fluorescence detection system, a detection system for simultaneously detecting GyVg1 and GyH1 is established. The detection system can simultaneously detect two different subtypes of gyrovirus, improves the detection efficiency, and provides a new technical means for the dynamic study of virus infection.

The present application relates to the field of virus detection technology, specifically disclosing a dual microfluidic fluorescent PCR detection kit for the H3N2 subtype of avian influenza virus. The dual microfluidic fluorescent PCR detection kit comprises a sample lysis extract for detecting the H3N2 subtype of avian influenza virus, a sample wash solution I, a sample wash solution II, and a reaction premix. The reaction premix contains three sets of specific sequence combinations: an H3-specific sequence combination, an N2-specific sequence combination, and an internal standard gene-specific sequence combination. By designing specific primers and probes, and performing optimization and screening, the present application enables simultaneous identification of H3 and N2 subtype AIVs in a single reaction. Furthermore, the amplification reactions of the two target genes are free of interference, resulting in high specificity, high sensitivity, and good reproducibility. This kit enables accurate diagnosis of viral infection and has important application value in the prevention and control of avian influenza epidemics.

The present invention belongs to the field of biotechnology and relates to anti-H10 subtype influenza virus hemagglutinin protein monoclonal antibody 2G9 and its application. Monoclonal antibody 2G9 is identified as IgG2a, κ type, and the application of the antibody is achieved through affinity purification, immunological methods and other technologies. Anti-H10 subtype influenza virus hemagglutinin protein monoclonal antibody 2G9, the monoclonal antibody subtype is IgG2a, κ type, named 2G9, can specifically recognize the hemagglutinin protein of H10 subtype influenza virus, the amino acid sequence of the heavy chain variable region of the antibody is shown in SEQ ID No. 2, and the amino acid sequence of the light chain variable region is shown in SEQ ID No. 4. The present invention provides an effective tool for the auxiliary diagnosis of H10 subtype influenza virus infection in clinical samples, and can be promoted and applied to various detection technologies as well as clinical and experimental research.

The application discloses a recombinant nanobody against swine influenza virus of H1N1 subtype, and a preparation method and application thereof, and belongs to the field of biological medicines. A VHH sequence of the nanobody against swine influenza virus of H1N1 subtype is screened from a nanobody bacterial library of an immunized llama, the VHH sequence is directionally cloned into a pPIC9K vector to construct a recombinant eukaryotic expression plasmid, and the recombinant eukaryotic expression plasmid is transformed into a Pichia pastoris expression system to screen an expression engineering strain, and the recombinant nanobody is obtained after induction culture of the expression engineering strain. It is proved by in-vitro and in-vivo experiments that the recombinant nanobody developed in the application can neutralize CA / 04 and has a remarkable therapeutic effect. Therefore, the neutralizing nanobody provided in the application has considerable potential in treating CA / 04. The findings will provide experimental data for developing nanobody preparations against swine influenza virus of H1 subtype and provide a new strategy for preventing and controlling swine influenza.

The invention belongs to the technical field of virus detection, and particularly relates to a primer composition for detecting an avian influenza NA gene, a detection method and a detection device. The avian influenza NA genes comprise NA genes of an influenza A virus H5N1, an influenza A virus H5N6, an influenza A virus H7N7, an influenza A virus H7N9 and an influenza A virus H9N2, and the primer composition comprises nucleotide sequences as shown in SEQ ID No.1-25. The primer composition for detecting the NA gene causing the highly pathogenic human and livestock co-infected avian influenza, provided by the invention, can be used for detecting whether a sample contains the NA gene causing the highly pathogenic human and livestock co-infected avian influenza, namely NA genes of influenza A virus subtypes H5N1, H5N6, H7N7, H7N9 and H9N2, and a result can be conveniently, quickly and accurately detected; in a practical application process, a detection result can be obtained within 40 minutes.

The present application belongs to the field of genetic engineering and biological medicine technology, and particularly relates to a biomaterial for regulating HTRA1 gene expression and application thereof in antiviral. Specifically, the present application finds that inhibiting HTRA1 gene can significantly promote replication of H13N2 subtype influenza virus, thereby promoting infection of H13N2 subtype influenza virus, while overexpression of HTRA1 can effectively inhibit replication of H13N2 subtype influenza virus, reduce virus titer, and thus improve the ability to resist influenza virus infection, so that the HTRA1 gene can be used as a key target for regulating replication of H13N2 subtype influenza virus, and thus has good practical application value.

The present invention discloses a 2.3.4.4b clade H5 subtype avian influenza virus strain and its application. The 2.3.4.4b clade H5 subtype avian influenza virus strain is avian influenza virus A / chicken / Shenyang / 12301 / 2024, which is deposited in the General Microbiological Center of the China Committee for Culture Collection of Microorganisms, with the deposit number of CGMCC No. 46299 and the deposit date of December 12, 2024. By searching for a 2.3.4.4b clade H5 subtype AIV with good cross-immune reaction as a vaccine candidate strain and preparing a whole virus inactivated vaccine, the present invention aims to effectively prevent and control the infection of recently prevalent H5 subtype AIV in poultry, and further eliminate the potential threat of influenza virus in poultry farming to public health safety.

The invention relates to the technical field of molecular biology. The invention provides a primer probe combination for detecting an H10 subtype avian influenza virus, the primer probe combination comprises a primer pair and a probe for detecting the H10 subtype avian influenza virus, and the primer pair for detecting the H10 subtype avian influenza virus comprises an H10-F sequence as shown in SEQ ID NO.1 and an H10-R sequence as shown in SEQ ID NO.2; and the probe for detecting the H10 subtype avian influenza virus is an H10-P sequence as shown in SEQ ID NO.3. The invention further discloses a kit for detecting the H10 subtype avian influenza virus. The primer probe combination disclosed by the invention can be used for detecting and distinguishing epidemic H10 viruses of European subpedigree and North American pedigree, does not generate cross reaction with other common viruses, and is high in detection sensitivity, and the minimum detectable quantity is 5 copies.

The application discloses a chicken Newcastle disease, avian influenza and chicken infectious rhinitis triple inactivated vaccine and preparation and application thereof, and relates to the technical field of bioengineering. The application discloses a preparation method and application of a chicken Newcastle disease, avian influenza and chicken infectious rhinitis (A, B and C three serotypes) subunit protein triple inactivated vaccine. The preparation method comprises the following steps: constructing and expressing a serum B paracolon avian bacillus subunit protein expression strain; and preparing the chicken Newcastle disease, H9 subtype avian influenza and chicken infectious rhinitis (A, B and C three serotypes) subunit protein triple inactivated vaccine at a certain ratio. The triple inactivated vaccine has good safety, can be used for the prevention of chicken Newcastle disease, H9 subtype avian influenza and A, B and C three serotypes paracolon avian bacillus diseases in a laying hen group, and does not cause the decrease of the laying rate after immunization. Meanwhile, the paracolon avian bacillus subunit protein can improve the antibody level of the Newcastle disease and the H9 avian influenza, and the protection of the vaccine on the A, B and C three serotypes paracolon avian bacillus can reach 100%.

The application discloses a kind of avian influenza and avian infectious bronchitis two-union recombinant subunit vaccine and its preparation method and application.The recombinant protein is by replacing the head domain of AIV HA protein with the RBD domain of IBV S protein, while adding GCN4, Trimer-tag trimer tag in sequence, the recombinant protein obtained, the recombinant protein is expressed in vitro using insect cells and prepared into subunit vaccine.The vaccine prepared by the application can produce high-level neutralizing antibodies against homologous and heterologous H9N2 subtype avian influenza virus after immunizing chicken population, while significantly reducing the virus discharge level and the histopathological changes caused, and can also induce high-level specific antibodies and neutralizing antibodies against QX subtype avian infectious bronchitis virus, effectively inhibit discharge, viral load, and reduce tissue lesions.The application does not depend on chicken embryo, and has low production cost and short cycle, and can simultaneously prevent avian influenza and avian infectious bronchitis.

The invention belongs to the technical field of gene engineering and biological medicine, and particularly relates to a biological material for inhibiting IDE gene expression and application of the biological material in anti-influenza virus. Researches find that specific inhibition of expression of the IDE gene can significantly inhibit infection of influenza viruses, especially H13N2 subtype influenza viruses. Specifically, the research finds that the mRNA expression level of the NP gene of the H13N2 subtype influenza virus in the A549 cell line for specifically inhibiting the IDE gene expression and the expression level of the PB2 protein are obviously reduced; meanwhile, the virus titer after the H13N2 subtype influenza virus infects the A549 cell line specifically inhibiting IDE gene expression is also obviously lower than that of a control group. The invention proves that the infection of the H13N2 subtype influenza virus can be obviously inhibited by specifically inhibiting the expression of the IDE gene, and a new thought is provided for resisting influenza virus infection, so that the method has a good practical application value.

The application belongs to the technical field of biology, and relates to an anti-H10 subtype influenza virus hemagglutinin protein monoclonal antibody and application thereof. The application is obtained by using cell engineering and antibody engineering technology, and is a hybridoma cell line secreting the anti-hemagglutinin protein monoclonal antibody. The anti-hemagglutinin protein monoclonal antibody 1E1 is prepared by inducing ascites of the same strain mouse, and is identified as IgG2a and kappa type. The application of the antibody is realized by affinity purification and immunization methods. The application can specifically recognize the hemagglutinin protein of the H10 subtype influenza virus. The heavy chain variable region amino acid sequence of the antibody is shown in SEQ ID No. 2, and the light chain variable region amino acid sequence is shown in SEQ ID No. 4. The application provides an effective tool for auxiliary diagnosis of H10 subtype influenza virus infection in clinical samples, and can be applied to various detection technologies and clinical and experimental researches.

The invention relates to the technical field of molecular detection, in particular to an RPA (recombinase polymerase amplification) primer, a probe and a detection method for detecting H9 subtype avian influenza virus. The RPA primer is divided into an upstream primer RPA-F and a downstream primer RPA-R, and the nucleotide sequence of the upstream primer RPA-F is any one as shown in SEQ ID NO.1-4; the nucleotide sequence of the downstream primer RPA-R is any one as shown in SEQ ID NO.5-8; the nucleotide sequence of the probe is as shown in SEQ ID NO. 9; wherein the RPA primer is designed on the basis of the HA gene sequence of the H9 subtype avian influenza virus, the specificity is relatively high, and the detection accuracy is high; the detection method has high detection sensitivity, isothermal amplification of the H9 subtype AIV can be completed at 39 DEG C within 20 min, and the problems that a conventional detection method needs long time, dependence on instruments and equipment is high, and the false positive result of the detection result is high are solved.

The present invention relates to a chimeric influenza virus hemagglutinin (HA) polypeptide comprising one or more stem domain sequences fused to one or more globular head domain sequences, each of the one or more stem domain sequences having at least 60% homology to a stem domain consensus sequence of an H1 subtype HA (H1 HA) and / or an H5 subtype HA (H5 HA), each of the one or more globular head domain sequences having at least 60% homology to a globular head domain consensus sequence of an H1 subtype HA (H1 HA) or an H5 subtype HA (H5 HA).

The invention belongs to the field of biology, and discloses preparation and application of a non-methanol-induced expression H5 subtype avian influenza virus subunit antigen and a vaccine thereof, PDAK is applied to pichia pastoris expression of avian influenza virus H5 subtype HA protein, an HA protein multiple-insertion recombinant strain with a PDAK promoter is successfully constructed, and in the research process, it is found that the H5 subtype avian influenza virus subunit antigen and the vaccine thereof have the advantages that the H5 subtype avian influenza virus subunit antigen and the vaccine thereof can be used for preparing the H5 subtype avian influenza virus subunit antigen; by adopting the recombinant plasmid disclosed by the invention, multiple insertions can be generated in a pichia pastoris strain, the target protein expression quantity can be obviously improved, and the foreign protein expression efficiency of the recombinant plasmid is superior to that of a PAOX1 promoter and a PGAP promoter; according to the invention, the HA protein of the PDAK promoter is multiply inserted into the recombinant strain to express the HA protein to prepare the subunit vaccine, after the vaccine is immunized, a higher hemagglutination inhibition antibody level can be generated aiming at homologous strains, and the survival rate of experimental chickens after challenge is 100%.