126 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 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 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 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 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 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 discloses a composite insecticide targeting aedes aegypti PGRP gene and application thereof, the insecticide is a preparation taking double-stranded RNA of a key PGRP gene as an effective component, and the double-stranded RNA comprises a gene aiming at PGRP-LB subtype and coding dsPGRP-LB and a gene aiming at PGRP-SC2 subtype and coding dsPGRP-SC2; wherein the nucleotide sequence of a front primer of the gene for coding the dsPGRP-LB is as shown in SEQ ID NO. 1; the nucleotide sequence of a rear primer for coding the gene dsPGRP-LB is as shown in SEQ ID NO. 2; the nucleotide sequence of a front primer for coding the gene dsPGRP-SC2 is as shown in SEQ ID NO. 3; and the nucleotide sequence of a rear primer of the gene for coding the dsPGRP-SC2 is as shown in SEQ ID NO. 4. According to the invention, the target specificity is strong, non-target biological interference is avoided, the insecticidal efficiency is obviously improved, the synergistic increasing agent has good suitability, mistaken killing of beneficial organisms in an ecological system is avoided, and accurate mosquito control is realized.

This invention provides a microfluidic chip for the multiplex differentiation and diagnosis of different avian influenza virus subtypes and its application. Specifically, it is a microfluidic chip based on TaqMan probes that can simultaneously and rapidly differentiate and diagnose 25 avian influenza subtypes, enabling high-throughput simultaneous detection of multiple gene targets. This invention utilizes high-throughput microfluidic chip technology to simplify the differential diagnosis procedure for avian influenza virus subtypes, resulting in more accurate and time-saving results. It is more suitable for rapid clinical diagnosis of avian influenza, providing technical support for rapid clinical detection, diagnosis, and prevention of avian influenza, and is of great significance for epidemic prevention and control and safeguarding public health security.

The application belongs to the field of biotechnology, and relates to preparation and application of an anti-H10 subtype influenza virus hemagglutinin protein neutralizing monoclonal antibody. The anti-H10 subtype influenza virus hemagglutinin protein neutralizing monoclonal antibody 1E10 is of IgG2b and kappa type, can be specifically combined with an H10 subtype influenza virus hemagglutinin protein antigen, 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 monoclonal antibody can effectively neutralize and treat H10 subtype influenza virus infection. The application provides an effective tool for clinical treatment of H10 subtype influenza virus infection, can be popularized and applied, and provides a new selection scheme for prevention and treatment of the anti-H10 subtype influenza virus.

The invention relates to a multi-antigen component PEI lipid delivery system and an anti-avian influenza virus multivalent vaccine comprising the same, belongs to the technical field of biological medicines, and provides a delivery system consisting of PEI, squalene, cholesterol and mPEG. The preparation method comprises the following steps of: constructing mRNA (messenger ribonucleic acid) molecules and a composite vaccine antigen system consisting of prokaryotic expression purified H7N9 avian influenza NP antigen, loading the composite vaccine antigen system into a delivery system to obtain the nanoparticles with a virus-like structure, and displaying the HA protein antigen and the NP antigen in the structure of the nanoparticles in a particle form by the nanoparticles, so that the nanoparticles can be used for preparing the H7N9 avian influenza NP / H7N9 avian influenza NP / H7N9 avian influenza NP / H7N9 avian influenza virus. And the mRNA molecule for coding the NA antigen, which is combined with the mRNA molecule, can be delivered into a cell in a manner of fusing lipid and a cell membrane, so that the mRNA molecule can be effectively expressed to induce a body to generate specific antibody reactions aiming at HA, NA and NP, and the body can also generate specific CTL reactions aiming at HA, NA and NP of different influenza virus strain subtypes.

This invention relates to the field of biology and discloses a nanobody sequence against H9 subtype avian influenza virus, its expression engineered strain, and preparation method. The invention screened a variable region sequence of a heavy chain antibody against H9 subtype avian influenza virus from an established VHH phage library immunized with alpacas. The nucleotide sequence is SEQ ID NO:1, and the encoded amino acid sequence is SEQ ID NO:2. This sequence was directionally cloned into the eukaryotic expression plasmid pPIC9K to construct a recombinant eukaryotic expression plasmid. This plasmid was then transformed into a Pichia pastoris expression system to screen for expression engineered strains. After methanol induction, supernatant collection, and affinity chromatography purification, the recombinant nanobody was obtained.

The invention discloses an RT-RAA-CRISPR / Cas13a visual detection kit for on-site rapid detection of a swine influenza virus and an application of the RT-RAA-CRISPR / Cas13a visual detection kit. The kit comprises a set of universal molecular detection composition for broad-spectrum detection of different subtype swine influenza viruses, the composition comprises a specific primer pair used for RT-RAA system amplification, a specific crRNA used for CRISPR / Cas13a system targeted recognition, and an RNA report probe used for carrying out amplification and visual output on a target recognition signal. Based on the sequence conservative property of each subtype M gene of the swine influenza virus, the kit can realize the broad-spectrum visual detection of the swine influenza virus on site. The method has the advantages of short detection time, simple operation, strong specificity and high sensitivity, and is expected to provide a new visual detection means for on-site rapid screening of swine influenza viruses.

The invention discloses a recombinant nano antibody for resisting H1N1 subtype A swine influenza virus as well as a preparation method and application thereof, and belongs to the field of biological medicines. The preparation method comprises the following steps: screening a VHH sequence for resisting an A H1N1 subtype swine influenza virus from a nano antibody bacterial library of an immune alpaca, directionally cloning the VHH sequence into a pPIC9K vector to construct a recombinant eukaryotic expression plasmid, transforming the recombinant eukaryotic expression plasmid into a pichia pastoris expression system to screen and express an engineering strain, and carrying out induced culture on the engineering strain to obtain the recombinant nano antibody. In-vitro and in-vivo experiments prove that the recombinant nano antibody developed by the invention can neutralize CA / 04 and shows a remarkable treatment effect. Therefore, the neutralizing nano antibody provided by the invention has great potential in the aspect of treatment of CA / 04. The discovery provides test data for research and development of anti-H1 subtype swine influenza virus nano antibody preparations, and provides a new strategy for prevention and control of swine influenza.

The invention belongs to the technical field of nano-antibodies, and particularly relates to a nano-antibody for resisting H1N1 influenza virus NA protein as well as a preparation method and application of the nano-antibody. The method comprises the following steps: by taking A / Puerto Rick / 8 / 34 (H1N1) influenza virus as a research object, purifying and inactivating the influenza virus through sucrose density gradient centrifugation, immunizing alpaca by taking the influenza virus as an immunogen, and screening by utilizing a phage display technology to obtain a high-affinity spectral anti-avian influenza virus NA protein nano antibody. Animal experiments show that the nano antibody has a good prevention effect when acting on A / Puerto Radio / 8 / 34 (H1N1) influenza virus infected C57 mice. Through preparation of the anti-influenza virus NA protein nano antibody, not only is a material provided for development of an H1N1 influenza virus diagnostic reagent, but also a thought is provided for identification of H1N1 subtype influenza virus epitope and vaccine design.