215 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 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 discloses a recombinant HVT for co-expressing an H9 subtype AIV HA gene, an IBV S gene and an IBDV VP2 gene and application of the recombinant HVT, and belongs to the technical field of medicine or veterinary medicine. The recombinant HVT is a recombinant HVT vaccine strain H30201 for co-expressing the H9 subtype AIV HA gene, the IBV S gene and the IBDV VP2 gene, which is obtained by inserting an expression cassette containing the H9 subtype AIV HA gene and the QX type IBV S gene between 95322-95323 nucleotides of an HVT FC126 strain and inserting an expression cassette containing the IBDV classic virulent strain VP2 gene between 112071-112088 nucleotides of an HVT genome at the same time through virus rescue. The invention also provides application of the recombinant HVT vaccine strain in preparation of vaccines for H9 subtype avian influenza, infectious bronchitis and infectious bursal disease.

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 belongs to the technical field of biology, and discloses a recombinant turkey herpesvirus co-expressing an avian influenza H9 gene and an avian adenovirus 4 type Fiber-2 gene, the H9 gene containing a promoter and a terminator is inserted between an HVT053 site and an HVT054 site of the turkey herpesvirus, the Fiber-2 gene containing a promoter and a terminator is inserted between an HVT087 site and an HVT088 site of the turkey herpesvirus, and the recombinant turkey herpesvirus co-expressing the avian influenza H9 gene and the avian adenovirus 4 type Fiber-2 gene is obtained. The recombinant virus is obtained. The recombinant turkey herpesvirus can stably co-express the H9N2 subtype avian influenza virus H9 protein and the fowl adenovirus 4 type Fiber-2 protein, has very excellent immune performance, and effectively improves the immune effect of the recombinant virus of the traditional fowl adenovirus Fiber-2 gene; compared with the recombinant herpesvirus of turkey which singly expresses any one of the H9N2 subtype avian influenza virus H9 protein and the fowl adenovirus 4 type Fiber-2 protein, the recombinant herpesvirus of turkey has the advantage that the performance of the recombinant herpesvirus of turkey is not weakened. Meanwhile, the invention also provides application and a vaccine of the recombinant herpesvirus of turkeys.

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 discloses a primer probe group for simultaneously detecting H5, H7 and H9 subtype and universal avian influenza viruses. The kit comprises a probe of the H5 subtype avian influenza virus, an upstream primer of the H5 subtype avian influenza virus, a downstream primer of the H5 subtype avian influenza virus, a probe of the H7 subtype avian influenza virus, an upstream primer of the H7 subtype avian influenza virus, a downstream primer of the H7 subtype avian influenza virus, a probe of the H9 subtype avian influenza virus and an upstream primer of the H9 subtype avian influenza virus. The kit comprises an upstream primer of the H9 subtype avian influenza virus, a downstream primer of the H9 subtype avian influenza virus, a probe of the universal avian influenza virus, an upstream primer of the universal avian influenza virus and a downstream primer of the universal avian influenza virus. The invention also discloses a kit containing the primer probe group. The kit has the advantages of being simple and convenient to operate, high in sensitivity, good in specificity, capable of detecting samples anytime and anywhere, low in cross contamination rate, rapid in result obtaining and the like, suspicious samples can be found and diagnosed in time, and the rapid detection capacity of the avian influenza virus is improved.

The invention relates to the technical field of molecular biology, in particular to a nucleic acid molecule composition for detecting H9 subtype avian influenza virus based on RT-RAA / CRISPR-Cas13a / LFD, a reagent, a kit and application. The nucleic acid molecule composition comprises an upstream primer RAA-H9-F, a downstream primer RAA-H9-R and a specific crRNA-H9, wherein the upstream primer RAA-H9-F is used for detecting the nucleic acid molecule; the sequence of the upstream primer RAA-H9-F is as shown in SEQ ID No: 1, the sequence of the downstream primer RAA-H9-R is as shown in SEQ ID No: 2, and the sequence of the specific crRNA-H9 is as shown in SEQ ID No: 3. The nucleic acid molecule composition provided by the invention can realize visual detection of the H9 subtype avian influenza virus, is high in detection accuracy, short in time consumption, good in specificity and high in sensitivity, provides a basis for rapid, simple and accurate diagnosis of the H9 subtype avian influenza virus, and meets the requirements of clinical detection.

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.

The invention relates to a heat-resistant H1N1 subtype influenza virus mutant vaccine strain and application thereof. The invention belongs to the technical field of virus gene engineering, and particularly discloses a heat-resistant H1N1 subtype influenza virus mutant vaccine strain and application thereof. According to the H1N1 subtype influenza virus mutant vaccine strain and the preparation method thereof, the HA protein head region, which is a key region influencing the virus thermal stability, is mutated, the virus thermal stability can be remarkably improved only by mutating three amino acid sites, meanwhile, the virus still has good thermal stability after being inactivated, the H1N1 subtype influenza virus mutant vaccine strain prepared by HA protein point mutation has the preservation number of CCTCC NO: V202471, and the H1N1 subtype influenza virus mutant vaccine strain can be used for preparing the H1N1 subtype influenza virus mutant vaccine strain. And immune animals can be induced to generate high-level HI antibodies.

The invention discloses a 2.3. 4.4b branch H5 subtype avian influenza virus strain and an application thereof. The 2.3. 4.4b branch H5 subtype avian influenza virus strain is an A type avian influenza virus A / chick / Shenyang / 12301 / 2024, is preserved in the China General Microbiological Culture Collection Center (CGMCC), has a preservation number of CGMCC No.46299, and is preserved for December 12, 2024, and the strain has a preservation number of CGMCC No.46299, a preservation number of CGMCC No.46299, a preservation number of CGMCC No.46299 and a preservation number of CGMCC No.46299. According to the invention, 2.3. 4.4b branch H5 subtype AIV with good cross immunoreaction is searched as a vaccine candidate strain to form a whole virus inactivated vaccine, so as to effectively prevent and control the infection of the latest epidemic H5 subtype AIV to poultry, and further eliminate the potential threat of influenza virus to public health safety in poultry breeding.

The invention discloses a veterinary vaccine composite adjuvant as well as a preparation method and application thereof. Active ingredients in the vaccine composite adjuvant are a manganese-containing compound and a Mincle agonist. According to the veterinary vaccine composite adjuvant disclosed by the invention, Mn and TDB are combined, so that the immunological enhancement effect of the TDB adjuvant is achieved. The composite adjuvant can induce a strong immune response reaction, significantly improves the immune effect, and is free of residual damage in use. Compared with the H7N9 vaccine prepared by singly using DDA-TDB, the H7N9 vaccine prepared by mixing the H7N9 subtype avian influenza inactivated virus as the antigen with the composite adjuvant can enhance humoral immunity and cellular immune response of a body, reduce virus load and improve protection level, and provides a product and technical support for production of veterinary vaccines.

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 relates to the technical field of molecular biology, in particular to a nucleic acid molecule composition for detecting H7 subtype avian influenza virus based on RT-RAA / CRISPR-Cas13a / LFD, a reagent, a kit and application. The nucleic acid molecule composition comprises an upstream primer RAA-H7-F, a downstream primer RAA-H7-R and a specific crRNA-H7, wherein the upstream primer RAA-H7-F and the downstream primer RAA-H7-R are used as primers of the nucleic acid molecule The sequence of the upstream primer RAA-H7-F is as shown in SEQ ID No: 1, the sequence of the downstream primer RAA-H7-R is as shown in SEQ ID No: 2, and the sequence of the specific crRNA-H7 is as shown in SEQ ID No: 3. The nucleic acid molecule composition provided by the invention can realize visual detection of the H7 subtype avian influenza virus, is high in detection accuracy, short in time consumption, good in specificity and high in sensitivity, provides a basis for rapid, simple and accurate diagnosis of the H7 subtype avian influenza virus, and meets the requirements of clinical detection.

The invention belongs to the technical field of biology, and relates to an anti-H6 subtype avian influenza virus hemagglutinin protein monoclonal antibody 1H1 and application thereof, cell engineering and antibody engineering technologies are utilized to obtain a hybridoma cell line for secreting a hemagglutinin monoclonal antibody, ascites is induced by mice of the same strain to prepare the anti-hemagglutinin protein monoclonal antibody 1H1, the anti-hemagglutinin protein monoclonal antibody 1H1 is identified as an IgG1, kappa type, and the anti-hemagglutinin protein monoclonal antibody 1H1 is identified as an anti-H6 subtype avian influenza virus hemagglutinin protein monoclonal antibody. And the application of the antibody is realized through technologies of affinity purification, immunization and the like. The monoclonal antibody has the advantages that the preparation method is simple and easy to implement, more importantly, the monoclonal antibody prepared by the method can have multiple purposes, an effective tool is provided for auxiliary diagnosis of H6 subtype avian influenza virus infection in a clinical sample, and the monoclonal antibody can be popularized and applied to multiple detection technologies and clinical and experimental research.

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.