315 results about "Highly pathogenic" patented technology

The application relates to a domesticated porcine reproductive and respiratory syndrome virus low virulent strain (JXA1-R strain), immunogenicity substance containing the viral strain, and a porcine reproductive and respiratory syndrome virus low virulent live vaccine developed through making use of the strain. The live vaccine is used to prevent the porcine reproductive and respiratory syndrome, in particular the highly pathogenic blue-ear porcine disease.

This invention discloses an nucleotide priemr tested by multiple real-time fluorescence RT-PCR, probe sequence and its method suitable for high-pathogenicity fowl-influenza H5, H7, H9 hypotype, relating to the nucleotide fragment RT-PCR augmenting primer and its complementary sequence, and the parting testing method of fowl-influenza H5, H7, H9 hypotype .

The invention discloses a virulent strain HuN4 for porcine reproductive and respiratory syndrome, and an attenuated vaccine strain HuN4-F112 (the collection number of microorganism: CGMCC No.2484) attenuated because of virulent strain HuN4 passage, and also discloses the application of the attenuated vaccine strain in preventing or curing the highly pathogenic blue-ear disease, which belongs to the biomedical field. The attenuated vaccine strain HuN4-F112 can be prepared into single-vaccine or combined strain (live vaccine or inactivated vaccine), can effectively prevent or cure the highly pathogenic blue-ear disease, and also can be prepared into diagnostic reagent for the highly pathogenic blue-ear disease diagnosis. The attenuated vaccine strain HuN4-F112 of the invention has the advantages of good security and high protection efficiency.

The invention provides a porcine reproductive and respiratory syndrome virus double-antibody sandwich ELISA kit. The kit comprises: an elisa plate coated with PRRSV N protein monoclonal antibody, an enzyme labeling PRRSV N protein monoclonal antibody, lysis solution and the like. A capture antibody and a detection antibody are respectively aimed at antigenic determinants with different N proteins.The kit provides a reliable means for quick detection of clinical PRRSV antigen. The kit can detects that blood serum only contains 0.2 TCID50 highly pathogenic PRRSV JXwn06 strain (non-highly pathogenic strain can be also be detected). Through detecting clinically collected 80 blood serum samples, compared with RT-PCR result, the specificity of the method is 88 percent, the sensitiveness is 90 percent and the coincidence rate of the specificity and the sensitiveness are 88.8 percent. The kit is convenient for operation, low in use cost, good repetitiveness and suitable for wide promotion andapplication.

The invention discloses a method for editing a large white pig CD163 gene by using CRISPR/Cas9. The large white pig CD163 gene is edited by using the CRISPR/Cas9, the extracellular domain SRCR5 of a CD163 receptor is destroyed, the 7th exon DNA fragment of the CD163 gene is knocked out, and the nucleotide sequence of the 7th exon of the CD163 gene is represented by SEQ ID NO.1. The edited gene obtained by adopting the method can completely resist infection of PRRSV including a highly pathogenic strain HP-PRRSV, allows the cell surface CD163 receptor expression to be normal, and has normal other biological functions.

The present invention relates to an artificially synthesized gene optiHA containing codons for chicken partial tropism. Its reading frame contains 1707 bp nucleotides and encodes a total of 568 amino acids. The gene is compatible with H5 subtype highly pathogenic avian influenza virus A/ Goose/GuangDong/1/96(H5N1)[GD/1/96(H5N1)]hemagglutinin (HA) gene has a nucleotide homology rate of 70%, an amino acid homology rate of 100%, and encodes the H5 subtype Hemagglutinin (HA) protein of avian influenza virus GD/1/96 (H5N1). The invention also relates to the application of the gene as an immunogenic gene of H5 subtype influenza DNA vaccine and other genetic engineering vaccines.

The invention discloses variant porcine reproductive and respiratory syndrome virus (PRRSV) TaqMan fluorescence quantitative RT-PCR detecting kit and application thereof. A primer and a TaqMan probe are designed and synthesized by referring to an NSP2 fragment gene sequence of the variant PRRSV and common PRRSV of a GenBank. By optimizing the reaction condition and constructing a standard plasmid product, a method for diagnosing the variant PRRSV by TaqMan fluorescence quantitative RT-PCR is established. A result indicates that the method has the advantages of strong specificity, high sensitivity, and the like and can detect the standard plasmid product with 264 copy numbers, and the virus quantity of 0.5623TICD50 is 10 times more sensitive than RT-PCR. By detecting 22 disease samples, 8 disease samples are positive, and the positive rate is 36.4 percent. Because the method has the advantages of quantification, high speed, accuracy, sensitivity, and the like, the invention is suitable for the diagnosis on the swinery infected variant PRRSV in the early stage, the medium stage and the later stage and plays an important role in effectively diagnosing, preventing and treating the highly pathogenic PRRSV.

The invention relates to a specific PRRS (Porcine Reproductive and Respiratory Syndrome) virus non-structural protein Nsp9 nanobody and discloses a VHH (Variable Domain of Heavy-chain Antibody) sequence of the nanobody and a DNA sequence encoding the nanobody at the same time. The invention further provides an expression vector which can express nanobody-eGFP fusion protein in an Marc145 cell. The Nsp9 nanobody can be specifically bound with PRRS virus non-structural protein Nsp9, and has functions of inhibiting multiplication of a classic strain and a highly pathogenic strain of a PRRS virus in the Marc-145 cell. The nanobody can be developed into a drug for treating a PRRS, and a gene corresponding to the nanobody can be used for development of a PRRS resistance transgenic pig.

The invention discloses a heat resisting protective agent of a bivalent live vaccine against highly pathogenic porcine reproductive and respiratory syndrome-pseudorabies, and a preparation method thereof. The heat resisting protective agent of the bivalent live vaccine against highly pathogenic porcine reproductive and respiratory syndrome-pseudorabies is prepared by the following materials by weight: 0.8 to 1.2% of gelatin, 4 to 6% of soybean peptone, 0.5 to 1.5% of glycine, 3 to 5% of sucrose, 2 to 3% of trehalose, 1 to 2% of polyvinyl pyrrolidone, 0.5 to 1% of mannitol, 0.1 to 0.2% of vitamin E and the rest being injection water. The preparation method comprises steps of high-temperature sterilization, filtration sterilization through a microporous membrane and mixing. The heat resisting protective agent of the bivalent live vaccine against highly pathogenic porcine reproductive and respiratory syndrome-pseudorabies of the invention reduces damage on virus activity caused by various physical and chemical factors during the freeze drying process, provides good protection and effectively solves the problems that a conventional protective agent of live vaccine needs refrigerated transport and storage is inconvenient.

The invention discloses a medicament for treating highly pathogenic porcine respiratory and reproductive syndrome and a method for preparing the same. The medicament is mainly prepared from rhubarb, dyers woad leaf, dandelion, southernwood, forsythia, gardenia, groundsel, plaster stone, scutellaria and astragalus. The preparation method comprises the following steps: according to a ratio, all materials are weighed up, the rhubarb is ground into powder, and soaked in boiling water for 30 minutes to form a leach solution, the 9 materials of dyers woad leaf, dandelion, southernwood, forsythia, gardenia, groundsel, plaster stone, scutellaria and astragalus are mixed for decoction and pentad temperature, the obtained liquid is mixed with the leach solution obtained after the rhubarb powder is soaked to prepare the medicament. The invention has the advantages of reasonable and unique formula, simple preparation process, low cost, little toxic and side effect and good control efficiency.

The invention discloses an influenza/human and avian influenza virus detection gene chip and a preparation method and an application thereof, 102 probes used for detecting A type, B type, H1N1 hipotype, H3N2 hipotype, H5N1 hipotype and H9N2 hipotype influenza viruses and 4 highly pathogenic differential diagnosis probes, as well as a process of sample treatment, hybridization, elution and chip identification which can carry out detection, typing or identification over the 6 types of viruses simultaneously and can identify the high pathogenicity of viruses. The invention not only greatly shortens the detection time of influenza viruses, but also improves the detection accuracy and provides a feasible technique support for the early warning mechanism of influenza epidemic situation in fields of clinical diagnosis, inspection quarantine, and the like.

The invention relates to a method for restraining enveloped virus infection, relative polypeptide and protein drug, belonging to biological medicine technical field. The invention comprises a method for restraining influenza virus, particularly for restraining highly pathogenic influenza virus and human influenza virus (as H1N1 hypotype and H3N2 hypotype), relative polypeptide and protein, relative nucleic acid encoding the polypeptide and protein, and relative carriers and cells for representing the polypeptide and protein.

The invention discloses a novel chicken infectious bursal disease virusVP 2c DNA (SEQ ID NO: 1), construction of its expression carrier, expressed recombined VP2 protein and the application of said recombined protein in preparing subunit genetic engineering vaccine against chicken infection bursal disease virus. The chicken infectious bursal disease virusVP 2c DNA can be highly expressed in yeast cell, and expressed recombined protein possesses biological activity and immunogenicity of the chicken infectious bursal disease virus natural protein . The expressed recombined protein can be produced into vaccine, and it is demonstrated through immunity chicken test that: the protein subunit vaccine can effectively induce body to generate spcial humoral immune response, make immunity chicken get 90% protection from deadly attack of highly pathogenic vv IBDV, and effectively prevent virus proliferation in the body.

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.

The invention relates to nucleic acid composition, a detection kit for an influenza virus and a micro-fluidic chip. The nucleic acid composition comprises detection primer pairs, wherein the detection primer pairs comprise at least two of the following primer pairs: an influenza A primer pair, an influenza B primer pair, an influenza C primer pair, an influenza A H1 primer pair, an influenza A H3 primer pair, an influenza A H5 primer pair, an influenza A H7 primer pair, an influenza A H9 primer pair, an influenza A N9 primer pair and a high-pathogenicity influenza A H7N9 primer pair. According to the nucleic acid composition, interference between the primers and probes can be avoided, at least two influenza viruses are detected simultaneously once, and detection sensitivity can reach 100 copies/mL.

The invention discloses an artificially cloned attenuated vaccine strains. The vaccine strains are strains cloned from attenuated vaccine strains HuN4-F112 of highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV), and marked restriction enzyme sites are introduced in the structural protein gene sequences of the strains. The invention also discloses a recombinant vector which comprises a full-length gene cDNA sequence of the attenuated vaccine strains HuN4-F112 of the highly pathogenic PRRSV. The 5'-end of the full-length gene cDNA sequence is additionally provided witha transcription promoter and the full-length gene cDNA sequence is internally introduced with the marked restriction enzyme sites. The artificially cloned attenuated vaccine strains of the invention can not only provide completely safe immune protection for resistance of an immune pig to the highly pathogenic PRRSV, but also effectively distinguish an immune pig of the PRRSV from a naturally infectious pig of the PRRSV, thus being beneficial to preventing and controlling the highly pathogenic PRRSV.

The invention relates to a heatproof lyophilized protective agent for a live vaccine JXA1-R strain for highly pathogenic porcine reproductive and respiratory syndrome and a method for preparing the same. The heatproof lyophilized protective agent for the live vaccine JXA1-R strain for the highly pathogenic porcine reproductive and respiratory syndrome in the invention is composed of the followingcomponents, by weigh: 0.8 to 1.2% of gelatin, 4 to 6% of soybean peptone, 0.5 to 1.5% of glycine, 4 to 6% of trehalose, 1 to 2% of polyvinyl pyrrolidone, 0.5 to 1% of mannitol, and the balance being water for injection; The heatproof lyophilized protective agent is prepared through the steps of high-temperature sterilization, microporous membrane filtration degerming and mixing. The heatproof lyophilized protective agent for the live vaccine JXA1-R strain for the highly pathogenic porcine reproductive and respiratory syndrome in the invention reduces damages to virus activity, the damages being caused by physicochemical factors in lyophilization of the vaccine, and enables virus loss rate to be lower before and after lyophilization with the virus loss rate being no more than 0.2 titer; further, the heatproof lyophilized protective agent satisfies the requirements for low-temperature refrigerated transport of the vaccine and effectively solves the problems such as inconvenient storage, high cost of the vaccine and the like.

Belonging to the field of traditional Chinese veterinary medicine, the invention discloses two traditional Chinese medicine compositions for treating highly pathogenic reproductive and respiratory syndrome. Wherein, one composition is mainly composed of crushed scutellaria, crushed andrographis paniculata, crushed dandelion, crushed Radix Astragali, crushed reed rhizome, crushed Herba Lopatheri,crushed herba fibraureae recisae, crushed honeysuckle, crushed forsythia and crushed licorice, and can be prepared into powder or premix. The other composition mainly consists of scutellaria extract, andrographis paniculata extract, dandelion extract, Radix Astragali extract, reed rhizome extract, Herba Lopatheri extract, herba fibraureae recisae extract, honeysuckle extract, forsythia extract and licorice extract, and can be prepared into injection, powder, aqueous agent, particulate agent or premix. The compositions of the invention have scientifically and reasonably proportioned materials, and the effective components can present increased curative effect through synergy. Thus, symptoms caused by highly pathogenic reproductive and respiratory syndrome can be alleviated and treated in different links. Able to address both the symptoms and root causes, the traditional Chinese medicine compositions of the invention have high cure rate, obvious curative effect, abundant medicine source, simple process, low cost, no medicine residue, and little toxic and side effect.

The invention relates to a porcine reproductive and respiratory syndrome virus RT-LAMP detection kit and a detection method thereof. Primers required by PRSSV RT-LAMP reaction system are designed according to the sequence of porcine reproductive and respiratory syndrome virus (PRSSV) published by GenBank; PRSSV virus RNA is extracted with the virus RNA extraction reagent (LBBII-RNA) designed and prepared by the inventor, the PRSSV RT-LAMP reaction system established in the invention is utilized for detection, and color developing agent is added after the reaction to judge the result; the result shows that the PRSSV virus RNA obtains efficient specific amplification after the reaction is conducted for 45 minutes at the temperature of 63 DEG C; and then, quick detection of porcine reproductive and respiratory syndrome virus American classical strain and NSP2 variant strain (highly pathogenic porcine reproductive and respiratory syndrome virus strain) is conducted by SpuI enzyme cutting. Compared with the prior art, the invention has quick detection, high sensitivity, low reaction cost, convenient and fast operation, which is capable of differentiating American classical strain and NSP2 variant strain (highly pathogenic porcine reproductive and respiratory syndrome virus strain) and meets the requirement of multi-level detection.

The invention discloses a recombinant porcine reproductive and respiratory syndrome virus as well as a preparation method and an application thereof. The recombinant porcine reproductive and respiratory syndrome virus provided by the invention is the recombinant virus which is obtained by carrying out substitution or deletion on a wild recombinant porcine reproductive and respiratory syndrome viral genome RNA; the substitution means carrying out substitution on all or a part of six structural proteins including a structural protein ORF2a, a structural protein ORF2b, a structural protein ORF3, a structural protein ORF4, a structural protein ORF5a and a structural protein ORF5 in the wild recombinant porcine reproductive and respiratory syndrome viral genome RNA as coding RNAs of labelled proteins; and the deletion means carrying out deletion on all or a part of coding RNAs of the six structural proteins in the wild recombinant porcine reproductive and respiratory syndrome viral genome RNA. The experiments prove that the recombinant virus has a good prevention effect on a high pathogenicity porcine reproductive and respiratory syndrome and has great importance to a porcine reproductive and respiratory syndrome viral vaccine.

The invention discloses ursolic acid saponin, a preparation method thereof and application in resisting highly pathogenic H5N1 influenza virus, and specifically relates to ursolic acid saponin compounds shown in general formula (I). A series of ursolic acid saponin is prepared from natural product ursolic acid serving as raw material by introducing an ester group into the C-28 site through structural modification at first, then introducing beta-glucosyl into the C-3 site, selectively protecting 3,6-OHs of the glucose by using BBTZ and introducing glycosyl into 2,4-OHs of the glucose. Pharmacological test shows that the ursolic acid saponin compounds have obvious inhibition effect on the invasion process of the H5N1 highly pathogenic influenza viruses on host cells and can be used as a medicament for preventing or treating the influenza viruses. The invention also discloses a pharmaceutical composition containing the ursolic acid saponin compounds and the combination of the compounds of the invention and other anti-virus medicines.

Transposon linker insertion mutagenesis of a full-length infectious clone of the highly pathogenic classical swine fever virus (CSFV) isolate Brescia (pBIC) was used to identify genetic determinants of CSFV virulence and host range. A virus mutant, RB-C22 (RB-C22v), possessing a 19-residue tag insertion at the carboxyl end of E1 was constructed. RB-C22v and the parental virus pBIC (pBICv) exhibited similar growth characteristics on primary porcine macrophage cell cultures although RB-C22v produced significantly smaller plaques on SK6 cell cultures. In vivo, RB-C22v was markedly attenuated in swine. In contrast with pBIC infection, where mortality was 100%, all RB-C22v-infected pigs survived infection remaining clinically normal. Additionally, chimeras of the Brescia strain and the attenuated vaccine strain CS were constructed and evaluated for viral virulence in swine. Chimeras 138.8v and 337.14v, chimeras containing the E2 glycoprotein of CS and chimeric virus 319.1v, which contained only the CS E2 glycoprotein in the Brescia background, were attenuated in swine. Chimeras encoding all Brescia structural proteins in a CS genetic background remained attenuated, indicating that additional mutations outside the structural region are important for CS vaccine virus attenuation. The combined results indicate a significant role for E1 glycoprotein and E2 glycoprotein in swine virulence.

The invention discloses reverse transcription-polymerase chain reaction (RT-PCR) detection for porcine reproductive and respiratory syndrome virus and classical swine fever virus and special primers for same. The invention provides the special primers for detecting the viruses. The special primers comprise a primer pair A and a primer pair B, wherein the primer pair A comprises a primer 1 and a primer 2; the primer pair B comprises a primer 3 and a primer 4; and nucleotide sequences of the primers 1, 2, 3 and 4 are sequences 1, 2, 3 and 4 in a sequence table. Experiments prove that: by the multiplex RT-PCR detection method for the porcine reproductive and respiratory syndrome virus and the classical swine fever virus, a classical porcine reproductive and respiratory syndrome virus strain, a high pathetic porcine reproductive and respiratory syndrome virus strain and the classical swine fever virus can be simultaneously detected through once reaction, and the method has a good application prospect on clinical diagnosis.