42 results about "Pathogenic avian influenza" patented technology

The invention belongs to the field of inspection and quarantine technology. Specifically, the invention is a nucleic acid detection reagent kit for synchronous discriminating and diagnosing avian influenza virus and newcastle disease virus. The detection reagents of the reagent kit include extraction reagent for extracting virus by silicon gel absorption column method, detection amplification reagent for detecting nucleic acid by RT-PCR Taq Man fluorescent probe method, and pretreatment liquid for solid tissue specimen for extracting virus RNA. Further, the invention employs in vitro transcription RNA as a positive contrast of the reagent kit. The reagent kit can rapidly and synchronously discriminate and diagnose avian influenza virus and newcastle disease virus which are highly infectious among avian plagues and have similar symptom, determine current major prevalent subtypes, such as H5, H7, H9, etc., and discriminate whether a infection source is an avian influenza having high pathogenicity, non-pathogenic avian influenza or mildly pathogenic avian influenza to human. The reagent kit is suitable for livestock and veterinarian station, import and export inspection and quarantine bureau, as well as other laboratories, and can be used for large-scale detection of influenza and epidemic surveillance.

Disclosed by the invention are an immunoassay kit and an immunoassay method for detecting highly pathogenic avian influenza virus subtype H5N1 rapidly, conveniently and specifically. Also disclosed are an immunochromatographic detection kit and an immunochromatographic detection method for detecting the virus subtype H5N1 rapidly, conveniently and specifically. It is found that a monoclonal antibody 4G6 produced by using the virus subtype H5N1 as an immunogen does not react with the subtype H5N2 virus or a subtype H5N3 virus and reacts only with a subtype H5N1 virus specifically. It is also found that only an avian influenza virus subtype H5N1 can be detected specifically by an immunoassay utilizing the monoclonal antibody 4G6. It is further found that the sensitivity of the detection of immunochromatography can be increased by adding a nonionic surface and a water-soluble vinyl polymer having a polar group containing an oxygen atom and a nitrogen atom to a developing solution to be used in the immunochromatography.

The invention relates to an scFv antibody for resisting H5N1 type highly-pathogenic avian influenza and an application thereof. A light chain amino acid sequence is shown as SEQIDNo: 1; a heavy chain amino acid sequence is shown as SEQIDNo: 2; a light chain nucleotide sequence is shown as SEQIDNo: 3; and a heavy chain nucleotide sequence is shown as SEQIDNo: 4. The scFv antibody for resisting the H5N1 type highly-pathogenic avian influenza is applied to the preparation of a medicine for resisting the H5N1 type avian influenza virus. Proven by protection tests of embryonated eggs, the 4F5 antibody has a 100% protective rate on prevention of the avian source and human source H5N1 type avian influenza virus infection, and the 4F5 antibody has a 100% protective rate on treatment of the avian source H5N1 type avian influenza virus infection. The highest protective rate on treatment of the human source H5N1 type highly-pathogenic avian influenza virus infection is 62.5%.

The invention discloses a recombinant H7N9 subtype avian influenza virus-like particle, and a preparation method and application thereof. Recombinant baculoviruses for expressing HA protein, NA protein and M1 protein of an H7N9 highly pathogenic avian influenza virus are respectively constructed on the basis of a recombinant baculovirus insect cell culture system. The three recombinant baculovirusstrains are used for co-infecting suspended insect cells, so that the H7N9 subtype avian influenza virus-like particle capable of being self-assembled in the cells can be obtained. After the concentration and purification with cane sugar with different gradient concentrations through an ultrafiltration tube, the H7N9 subtype avian influenza virus-like particle is mixed and emulsified with an adjuvant to prepare a vaccine. When a chicken is immunized by the prepared vaccine, the body can be induced to generate a specific antibody; and the advantages of high immunogenicity, good safety, high hereditary stability and the like are realized. After the attack by a lethal dose of H7N9 subtype highly pathogenic avian influenza viruses, the complete clinical protection can be provided, and virus expelling of the chicken is obviously inhibited. The invention provides a novel method for preventing H7N9 subtype avian influenza virus infection, and also lays a foundation for the development of a novel influenza virus vaccine.

A biochip for detecting avain fluenza virus of both human and fowls is prepared as providing activated chip substrate, providing various fluenza virus subset sections to be sample-set, providing quality control protein, carrying out arrangement pattern design of point measurement, making sample-sets of quality control protein and various avain fluenza virus subset sections according to prepared pattern for detecting out antibodies generated by various subsets simultaneously.

The invention relates to a kit for detecting multiple influenza viruses by a polymerase chain reaction (PCR) microarray. The kit comprises mainly a PCR microarray reaction plate, a PCR reaction solution, and a packaging box for centralized and partitioned packaging of reagent bottles or reagent tubes. Through the combination of a real-time fluorescence PCR technology and a microarray technology, on the single PCR microarray reaction plate, the kit can detect eight influenza viruses comprising influenza A virus, influenza B virus, seasonal influenza H1 subtype virus, seasonal influenza H3 subtype virus, influenza A H1N1 virus, highly pathogenic avian influenza H5 subtype virus, highly pathogenic avian influenza H7 subtype virus, and highly pathogenic avian influenza H9 subtype virus simultaneously. A result of detection adopting the kit can be utilized for differential diagnosis and monitoring of pathogens capable of causing influenza.

The invention discloses a highly-pathogenic avian influenza virus hemagglutinin protein identification method, which can be used for highly-pathogenic avian influenza virus hemagglutinin protein identification and provide a reference for understanding the structural characteristics of the highly-pathogenic avian influenza virus hemagglutinin protein; through the identification of the hemagglutinin protein, the method can indirectly distinguish the degree of the pathogenicity of the avian influenza virus strain to which the hemagglutinin protein belongs. The method comprises the following steps: a), to establish an amino acid holographic topological structure score vector, based on principal component analysis; b), to apply the amino acid holographic topological structure score vector to characterize the structure of the influenza virus hemagglutinin protein; c), to carry out normalized processing to the characteristic variable of each influenza virus hemagglutinin protein through a self-cross-covariance method; d), to establish an avian influenza virus hemagglutinin protein identification model through an RBF kernel-based support vector machine; e), to establish a highly-pathogenic avian influenza virus hemagglutinin protein identification model through the RBF kernel-based support vector machine.

The invention relates to recombinational bacillus subtilis(B.S.-HA) of expressing highly pathogenic avian influenza(AIV) HSN1 hemagglutinin HA protein, and belongs to the field of biotechnology genetic engineering. The recombinational bacillus subtilis(B.S.-HA) of expressing the highly pathogenic avian influenza(AIV) HSN1 hemagglutinin HA protein successfully establishes recombinational bacillus subtilis expression plasmid pHT43 of the highly pathogenic avian influenza hemagglutinin HA protein, and successfully conducts elec-transformation of the recombinational bacillus subtilis expression plasmid pHT43 of the highly pathogenic avian influenza hemagglutinin HA protein into bacillus subtilis WB800N bacterial strain (B.S.). As verified by Western-blot, the highly pathogenic avian influenza hemagglutinin HA protein can be successfully expressed in the recombinational bacillus subtilis. Oral immunization can stimulate chicken body of 10-day old chicken to generate an effective AIV specific immune response level. The recombinational bacillus subtilis(B.S.-HA) is hopeful to be developed to genetic engineering oral vaccine in preventing the highly pathogenic avian influenza.

The invention provides a polypeptide drug for treating diseases initiated by highly pathogenic bird flu virus H5N1. The polypeptide drug is characterized in that the amino acid sequence of the polypeptide drug is amino terminal-FLNVEWSYI-carboxyl terminal; a plurality of the amino acid sequences form a multicopy serial structure by using lysine as a joint. The polypeptide drug has the advantages of convenient transportation and storage and automatic batch production, and has the capability of activating specificity cytotoxicity T lymphocyte (CTL). The specificity CTL has a target cell which kills and dissolves H5 albumen with the highly pathogenic bird flu virus H5N1 with obvious effect without toxic side effect.

The invention relates to recombinant bacillus subtilis for expressing avian influenza virus HA antigen fragment and PB1-F2 protein in tandem, and belongs to the field of biotechnical genetic engineering. The invention successfully establishes a recombinant bacillus subtilis integrated expression plasmid plnHAPB1-F2 for expressing the avian influenza virus HA-PB1-F2 protein, and successfully achieves chemical conversion and electric shock conversion into bacillus subtilis WB600. Western-blot verification shows that the avian influenza virus HA antigen fragment and the PB1-F2 protein which are expressed in tandem can be successfully expressed in the recombinant bacillus subtilis. After one-day-old SPF chick is immunized through oral administration, a chick body can be stimulated to generate an effective HA-PB1-F2 specific immune response level, and nonspecific immunity is significantly enhanced. The recombinant bacillus subtilis is expected to be successfully developed into a genetic engineering oral vaccine for preventing the highly pathogenic avian influenza of poultry.

The invention relates to a polypeptide drug for treating diseases caused by highly pathogenic avian influenza virus H5N1, in particular to a polypeptide drug about diseases caused by highly pathogenic avian influenza virus H5N1, belonging to the field of biomedicine. The polypeptide drug is characterized in that the amino acid sequence of the polypeptide drug is amido-FLNVEWSYI-carboxyl, which is a special connection mode formed by using the characteristic that lysine has two amidoes and one carboxyl and using the lysine as a joint. The invention has the advantages of convenience for transportation and preservation and automated batch production; the polypeptide drug has the capacity of activating specific cytotoxicity T lymphocyte (CTL); and the specific CTL cells have target cells for killing and dissolving H5 protein having the highly pathogenic avian influenza virus H5N1, with obvious effects but without toxic or side effects.

The invention discloses a polypeptide vaccine for treating diseases caused by highly pathogenic avian influenza viruses H5N1 and relates to the field of biomedicine, in particular to a polypeptide vaccine for treating diseases caused by the highly pathogenic avian influenza viruses H5N1. The polypeptide vaccine is characterized in that the amino acid sequence of the polypeptide vaccine is amino-QIGNISWV-carboxyl; and the polypeptide vaccine forms a special connecting mode by utilizing the characteristics that the lysine has two amino groups and a carboxyl group and using the lysine as a joint. The polypeptide vaccine has the advantages of convenience for transportation and storage and automatic batch production and the capacity of activating specific cytotoxic T lymphocyte (CTL), wherein the specific CTL has the capacities of killing and dissolving target cells with the highly pathogenic avian influenza virus H5N1 protein, and the polypeptide vaccine has obvious effect and no toxic and side effects.

The invention relates to a new application of sakuranetin and derivatives thereof in preparation of an anti-highly pathogenic avian influenza (HPAI) H5N1 virus medicament. Experiments prove that the sakuranetin and the derivatives thereof have a good inhibiting and inactivating effect. The anti-H5N1 virus medicament can be prepared by using the compounds as an active ingredient. The anti-virus medicament can be used for clinically preventing and treating influenza viruses, particularly diseases caused by HPAI H5N1 viruses. The compounds are safe, low in toxicity and strong in pharmacologic action; and the raw material sources are rich, wide and cheap, the preparation cost is low, the process is simple, and the yield is high. In order to prevent and treat the influenza viruses, particularly the HPAI H5N1 viruses, the invention provides a new medicament source, which has great practical significance and a broad application prospect in the fields of medicine and bio-pharmaceuticals, particularly in the field of preparation of anti-H5N1 virus medicaments.

The invention discloses an avian influenza virus-like particle antigen, a vaccine, a preparation method of the avian influenza virus-like particle antigen and application of the avian influenza virus-like particle antigen or the vaccine. A strain of recombinant baculovirus based on tandem expression of HA, NA and M1 genes of an H7N9 subtype avian influenza virus transfects insect cells and performs self-assembly to form the avian influenza virus-like particle antigen; and the virus-like particle antigen prepared according to the invention is used for preparing the vaccine. After a chicken group is immunized by the vaccine prepared according to the invention, HI antibodies, neutralizing antibodies and IgY antibodies with the higher titer can be generated, 100-percent clinical protection onthe H7N9 subtype highly pathogenic avian influenza virus can be provided, and virus expelling can be effectively inhibited. The rely on the chicken embryos is avoided; the production cost is low; theperiod is short; and higher protection capability can be induced only by once low-dose immunization. A foundation is laid for industrial production of VLP vaccines.