29 results about "Types virus" patented technology

The utility model provides a pressure maintaining type virus removal filter which is characterized in that elastic membrane mounting seats are connected in a liquid inlet pipe and a liquid outlet pipe of the filter in a sealing manner, elastic membranes are assembled on the elastic membrane mounting seats, and cross-shaped openings are formed in the elastic membranes; the elastic membrane comprises an extrusion part, a connecting part and a convex part, an annular clamping groove is formed in the elastic membrane mounting seat, the extrusion part is tightly clamped in the annular clamping groove, the cross-shaped opening is formed in the convex part, the convex part protrudes towards the direction of the filter, and the opening and closing of the cross-shaped opening are controlled through the internal and external pressure difference of the elastic membrane so as to ensure the internal pressure of the filter. According to the pressure maintaining type virus removal filter, the elastic membranes are arranged in the liquid inlet pipe and the liquid outlet pipe, opening and closing of the cross-shaped opening are controlled through the internal and external pressure difference of the elastic membranes, when the pressure is interrupted, the cross-shaped opening of the elastic membranes is closed, liquid cannot pass through the cross-shaped opening, and certain pressure is kept in the filter; therefore, the virus penetration risk caused by pressure interruption is reduced.

This invention belongs to the field of genetic engineering. By introducing a TC tag between amino acids 199 and 200 of the wild-type bluetongue virus non-structural protein NS2, a recombinant virus was rescued. Analysis of viral plaques and growth curves revealed that the recombinant virus formed plaques of similar size to the wild-type virus, and their growth curves showed no significant difference. Immunofluorescence and FlasH-EDT2 staining of NS2 showed that the fluorescence of NS2 labeled with anti-NS2 antibody overlapped with that labeled with FlasH-EDT2. Three-dimensional imaging of FlasH-EDT2-labeled NS2 revealed that the viral inclusion bodies exhibited fluorescence around the periphery but no fluorescence inside, displaying an overall "core-shell" hierarchical structure. This recombinant bluetongue virus can be used for tracing and localizing NS2 protein and viral inclusion bodies in living cells, and can also be applied to BTV neutralization assays, antiviral drug screening, and other related research.

The present disclosure relates to a viral vector having an exogenous RNA segment having a hairpin-like structure, for example, having two or more base pairing regions separated by one or more non-base pairing regions. The exogenous RNA segment may have a secondary structure, minimum free energy, average position entropy, or other properties within a specified range, or have a value similar to one or more hairpin-like structures of a reference wild-type virus. In some examples, the viral vector is a live attenuated vaccine. In some examples, the viral vectors down-regulate a susceptibility gene in a host plant.

The present invention is related to method of preparing a gain-of-function mutation in poxvirus, like buffalopox virus (BPXV) by long-term propagating it in the presence of methyltransferase inhibitor like 3-Deazaneplanocin A (DZNep), which results in generation of viral 2'O-MTAse (VP39) mutant with a single point mutation (M236I). The resultant mutant has the following characteristics: (a) M236I mutation of 2'O-MTAse increases the cap-1 markings on viral mRNA; (b) The 2'O-MTAse mutant virus had hyperactive 2'O-MTAse and exhibited enhanced stability and translational efficiency of its mRNA; (c) The capping (cap1) performed by the mutant viral 2'O-MTAse (VP39) results in higher viral protein production, as compared to the wild type VP39 and (d) Mutant virus replicated at higher viral titer than the wild-type virus.

This invention discloses an attenuated chikungunya virus expressing a nanoluciferase gene, the nucleotide sequence of which is shown in SEQ ID NO:1. It is constructed by attenuating the virulence of the chikungunya virus LR2006-OPY1 strain by deleting 61 amino acids from positions 323 to 383 of the nsP3 protein and inserting a nanoluciferase gene after position 490 of the nsP3 protein. This virus model maintains similar growth characteristics to the wild-type virus but with significantly improved safety. Viral replication can be monitored in real time by detecting luciferase activity, making it suitable for drug screening at both cellular and animal levels. This provides an efficient and safe technical platform for the development of antiviral drugs against chikungunya virus.

The present invention relates to a composition for treating rhinovirus infectious diseases, comprising a GPR40 agonist as an active ingredient. Viruses causing cold symptoms include coronaviruses, picomaviruses, rhinoviruses, coxsackieviruses, adenoviruses, parainfluenza viruses, respiratory syncytial viruses, enteroviruses and the like, but since there is no universal antiviral agent capable of treating all types of viruses, application of the most appropriate therapeutic agent should vary depending on the virus causing the cold symptoms. The GPR40 agonist provided in the present invention exhibits a remarkable therapeutic effect on cold diseases caused by rhinovirus infection, and thus is expected to be actively used in the medical and healthcare fields.

Provided are an oncolytic virus preparation capable of crossing a blood-brain barrier, a preparation method therefor and a use thereof. The oncolytic virus preparation is an oncolytic virus modified by a polypeptide having a function of crossing a blood-brain barrier. The polypeptide having the function of crossing the blood-brain barrier is RVG29-Cys. The oncolytic virus is a genetically engineered oncolytic virus OH2 in which neurotoxic and immunosuppressive genes in a wild-type virus genome are knocked out and an immune-enhancing human granulocyte-macrophage colony stimulating factor gene is inserted. An OH2-PEG-RVG nanosystem is constructed by conjugating NHS-PEG-Mal to OH2, and then coupling a Mal group on NHS-PEG-Mal to a Cys group on RVG29-Cys, thus increasing the aqueous solubility of a drug while facilitating the therapeutic drug crossing of the blood-brain barrier to treat brain tumors, and preventing neutralizing antibodies and macrophages in vivo from neutralizing and phagocytosing the oncolytic virus, thereby prolonging the circulation time in vivo.

The invention relates to the technical field of viruses, in particular to a construction method and application of a porcine parvovirus full-length infectious clone carrying a stable genetic marker. The method comprises the following steps: cloning a complete genome of a porcine parvovirus SXND18 strain to a plasmid pBluescript II SK (+) through an In-fusion seamless cloning technology, so as to obtain a recombinant plasmid pSXND18; meanwhile, a new EcoRI restriction enzyme cutting site is introduced into a genome by utilizing an overlapping PCR (Polymerase Chain Reaction) technology and is used as a genetic marker for identifying a wild virus and a rescue virus; and mixing the recombinant plasmid with a transfection reagent, and transfecting ST cells to obtain the rescue virus rSXND18. The porcine parvovirus reverse genetic operating system established by the invention can be used for research in the directions of porcine parvovirus virulence analysis, cross-species transmission mechanism and the like, and also lays a foundation for research on novel vaccines of the porcine parvovirus.

This invention discloses PCR primers and probes for detecting ILT live vaccine virus and wild-type virus. The primers include a universal upstream primer: 5'-GCACAAAGTCCAAATCTTCT-3' and a universal downstream primer: 5'-TCAATCACAGACGGTACTTTC-3'. The probes include a vaccine virus-specific probe: 5'-FAM-CAAGTTGATGAATATGCTCAC-MGB-3' and a wild-type virus-specific probe: 5'-ROX-CAAGTTGATGAATGTGCTCA-MGB-3'. The 5' end of the vaccine virus probe is labeled with the FAM fluorescent group, and the 5' end of the wild-type virus probe is labeled with the ROX fluorescent group. The 3' ends of both probes are labeled with the MGB quencher group. Using these primers and probes, ILT live vaccine virus and wild-type virus can be detected and distinguished rapidly and accurately, and can be used for epidemic monitoring in farms.

The invention belongs to the technical field of biology, and particularly relates to an S1 protein trimer subunit vaccine of a high-immunogenicity G2c subtype porcine epidemic diarrhea virus (PEDV) strain obtained based on screening as well as a preparation method and application of the S1 protein trimer subunit vaccine. The effective component is a recombinant S1 protein trimer which is expressed in a CHO system by introducing an optimized I-type virus fusion protein HR2 trimerization structural domain and a His tag on the basis of a G2c subtype strain (HBXZ2404-19 strain) S1 gene sequence obtained on the basis of screening. Results prove that the prepared S1 trimer subunit vaccine not only can efficiently neutralize various same subtype G2c strains, but also can generate a remarkable neutralizing effect on other G2a subtype strains and G2b subtype strains, and the S1 trimer subunit vaccine is proved to have broad-spectrum cross protection capability.

The invention is applicable to the technical field of genetic engineering, and provides an E2-1b + mi3 protein nanoparticle of bovine viral diarrhea virus as well as a preparation method and application of the E2-1b + mi3 protein nanoparticle, the E2-1b + mi3 protein nanoparticle is formed by coupling an E2-1b-st fusion protein and a mi3-sc fusion protein, the 1b type E2 antigen of the bovine viral diarrhea virus is displayed on the surface of the mi3 protein nanoparticle through an isopeptide bond; the amino acid sequence of the mi3 protein nanoparticle is shown as SEQ ID NO: 6 in a sequence table; the amino acid sequence of the E2-1b-st fusion protein is as shown in SEQ ID NO: 2 in a sequence table; the amino acid sequence of the mi3-sc fusion protein is shown as SEQ ID NO: 4 in a sequence table. After animals are immunized by the E2-1b + mi3 protein nano-particles, an effective immune protection effect can be generated, and the E2-1b + mi3 protein nano-particles have a good prevention and control application prospect on bovine viral diarrhea caused by BVDV 1b type viruses.

The invention relates to the technical field of microorganisms, in particular to a highly pathogenic duck virus hepatitis III virus and an egg yolk antibody, preparation and application thereof. The virus strain is preserved in the China General Microbiological Culture Collection Center (CGMCC) on December 24, 2024, the preservation number is CGMCC No.46271, and the virus strain is named as RD240820B. The egg yolk antibody is prepared by using the highly pathogenic duck virus hepatitis III type virus strain as an antigen. The egg yolk antibody is applied to preparation of products for preventing or treating duck virus hepatitis III. In a virulence experiment on a duck embryo, the death rate of the virus strain reaches 80% after the duck embryo is inoculated for 36 hours, and the virus strain has strong pathogenicity. And the death rate within 60 hours after inoculation is 100%, and the strain has relatively strong pathogenicity. The death rate is still 60% when the diluent is 10-8, and the toxicity is strong. The ELD50 value of the strain is relatively low, which indicates that the strain has relatively strong pathogenicity to duck embryos. After the egg yolk antibody is injected, the protection rate reaches up to 97.6% after the diseased duck is treated, and the egg yolk antibody has a remarkable prevention and control effect.

The invention belongs to the technical field of biology, and particularly discloses a cell line capable of stably expressing an nsP4 gene and application of the cell line in preparation of a replication-deficient chikungunya virus. The inventor uses retrovirus for expressing CHIKV nsP4 to infect hamster kidney cells (hamster kidney cells), after puromycin screening, verification of passage stability of monoclonal cells, protein expression stability, replication-deficient virus titer and the like is carried out, finally, a cell line which can be used for continuous passage of CHIKV replication-deficient viruses and has high titer is obtained, and the preservation number is CCTCC NO: C2025281. The replication-deficient virus CHIKV-delta nsP4 generated by the RNA of the nsP4 gene deleted from the trans-complementary CHIKV of the cell line has good safety, immunogenicity and immune protection efficacy, can effectively resist the challenge infection of wild viruses, and can be used for preparing a safe and effective CHIKV novel vaccine candidate strain.

This invention belongs to the field of viral genetic engineering technology and discloses a bovine herpesvirus type 1 (BHV-1) with three gene deletions, its construction method, and its applications. Using a Fosmid library, four myxovirus rescue combinations were screened. FosBHV-1-b-ΔTK-mCherry and FosBHV-1-d-ΔgEct / US9-eGFP were obtained through Red / ET and Gateway methods. These were then combined with FosBHV-1-a and FosBHV-1-c to construct an infectious cloning platform for recombinant BHV-1 with the TK / gEct / US9 three-gene deletions, rescuing recombinant viruses carrying tracer genes. This method has high recombination efficiency, and the cloning platform can efficiently construct BHV-1 gene-deleted strains. The immunogenicity of the deleted strains is comparable to that of wild-type viruses, with good safety, providing an efficient technical means for the development of BHV-1 attenuated vaccines and bovine disease live vector vaccines.

The present application relates to an attenuated oncolytic virus strain, an oncolytic virus vaccine and a drug for treating tumors by combining the oncolytic virus vaccine with immune cells. The present application provides a new attenuated oncolytic virus strain by a site-directed mutation of a matrix protein M of a VSV wild-type virus. On the basis of the attenuated oncolytic virus strain, the present application further provides a vaccine that can be used in tumor treatment. On the basis of the vaccine, the present application further provide a drug that can effectively treat multiple kinds of tumors by combining the vaccine with immune cells.

The invention discloses a totally-enclosed avian influenza H5, H7, H9 and universal virus quadruple detection kit. The kit comprises primers and probes of a universal avian influenza virus M gene, an H5 subtype avian influenza virus HA gene, an H7 subtype avian influenza virus HA gene, an H9 subtype avian influenza virus HA gene and a reference gene LconR. According to the invention, the ligand-dependent nuclear receptor corepressor protein is introduced as a reference gene, and the gene widely exists in mammals and birds and is very suitable for detection of multi-species infection epidemic diseases such as avian influenza. The internal reference can participate in nucleic acid extraction and amplification processes, strictly controls the quality of each detection step, fundamentally avoids false negative results, and significantly improves the reliability of detection results. The internal reference also expands the species application range of the detection method, so that avian influenza detection is not limited to poultry any more, and the method can be widely applied to all mammals possibly infected with avian influenza.

The invention is applicable to the technical field of genetic engineering, and provides E2-1b-mi3 fusion protein of bovine viral diarrhea virus as well as a preparation method and application of the E2-1b-mi3 fusion protein, and the E2-1b-mi3 fusion protein is formed by fusing a mi3 tag sequence with a 1b type E2 genome of the bovine viral diarrhea virus; the amino acid sequence of the mi3 tag sequence is as shown in a sequence table SEQ ID NO: 4; the amino acid sequence of the E2-1b-mi3 fusion protein is shown as SEQ ID NO: 2 in a sequence table. After animals are immunized by the E2-1b-mi3 fusion protein, an effective immune protection effect can be generated, and the E2-1b-mi3 fusion protein has a good prevention and control application prospect on bovine viral diarrhea caused by the BVDV 1b type virus.

The present invention relates to recombinant HVT vectors expressing antigens of avian pathogens and uses thereof. In particular, the present invention provides recombinant herpesvirus comprising and expressing a turkey (HVT) vector of an antigen of an avian pathogen, compositions comprising the recombinant HVT vectors, multivalent vaccines comprising the recombinant HVT vectors and one or more wild-type viruses or recombinant vectors. The present invention also provides methods of vaccinating against multiple avian pathogens and methods of making the recombinant HVT vectors.

The invention belongs to the technical field of biological detection, and relates to a probe primer combination for detecting AdV wild type virus residues and a method and application thereof. According to the invention, two sets of primer probe combinations respectively targeting wild type adenovirus E1A-CR2 and E3 regions are designed, and dPCR is utilized to realize dual-target synchronous detection and signal distinguishing. By calculating the concentrations of linkages of two targets on the same DNA chain, false positive results caused by host cell DNA residues, incomplete E3 region knockout or single region non-specific amplification can be effectively eliminated, so that the specific recognition of the wild type adenovirus is realized. The method is convenient and rapid, can complete detection of AdV wild virus residues in a short time, and does not need expensive instruments and equipment; compared with a PCR (Polymerase Chain Reaction) method (about 1 / 107), the method has the advantages that the sensitivity is improved by 2 orders of magnitude, and the sensitivity can be improved to 0.5 / 109 or above.

This invention belongs to the field of biodetection technology and relates to a method for specifically detecting wild-type virus residues in modified viral products and its application. This invention significantly suppresses non-specific extension into a mutant background by using LNA-modified primers; simultaneously, the SNP-embedded probe design differentiates at the signal level, amplifying the fluorescence signal differences between wild-type and mutant sequences. Finally, the ddPCR platform effectively eliminates PCR inhibitors and background noise from non-specific amplification. These three factors synergistically produce a significant enhancement effect. Taking VSV as an example, the embodiments demonstrate that this method can stably and repeatedly detect wild-type virus with a proportion as low as 1 / (10^7) against a mutant virus background of up to 10^10 copies, achieving a sensitivity at the part-in-ten-millions level. This represents an improvement of at least four orders of magnitude compared to traditional methods, far exceeding the reasonable expectations of those skilled in the art based on conventional technological improvements.

The invention is suitable for the technical field of gene engineering, and provides gD + mi3 protein nanoparticles for preventing and treating IBR and a preparation method and application thereof.The gD + mi3 protein nanoparticles are formed by coupling gD-st fusion protein and mi3-sc fusion protein, and gD antigens of infectious bovine rhinotracheitis viruses are displayed on the surfaces of the mi3 protein nanoparticles through isopeptide bonds; the amino acid sequence of the mi3 protein nanoparticle is shown as SEQ ID NO: 6 in a sequence table; the amino acid sequence of the gD-st fusion protein is as shown in a sequence table SEQ ID NO: 2; the amino acid sequence of the mi3-sc fusion protein is shown as SEQ ID NO: 4 in a sequence table. After animals are immunized by the gD + mi3 protein nanoparticles, an effective immune protection effect can be generated, and the gD + mi3 protein nanoparticles have a good application prospect in prevention and control of infectious bovine rhinotracheitis caused by IBRV (infectious bovine rhinotracheitis virus).

The invention is applicable to the technical field of genetic engineering, and provides an E2-1d + mi3 protein nanoparticle of bovine viral diarrhea virus as well as a preparation method and application of the E2-1d + mi3 protein nanoparticle, the E2-1d + mi3 protein nanoparticle is formed by coupling an E2-1d-st fusion protein and a mi3-sc fusion protein, the 1d type E2 antigen of the bovine viral diarrhea virus is displayed on the surface of the mi3 protein nanoparticle through an isopeptide bond; the amino acid sequence of the mi3 protein nanoparticle is shown as SEQ ID NO: 6 in a sequence table; the amino acid sequence of the E2-1d-st fusion protein is as shown in SEQ ID NO: 2 in a sequence table; the amino acid sequence of the mi3-sc fusion protein is shown as SEQ ID NO: 4 in a sequence table. After animals are immunized by the E2-1d + mi3 protein nano-particles, an effective immune protection effect can be generated, and the E2-1d + mi3 protein nano-particles have a good prevention and control application prospect on bovine viral diarrhea caused by BVDV 1d type viruses.

The invention relates to the technical field of animal infectious disease diagnosis and vaccines, in particular to a Brucella disease DIVA diagnosis antigen BruAb20955, and the amino acid sequence of the BruAb20955 protein is as shown in SEQ ID NO: 2. A nucleotide sequence for coding the brucellosis DIVA diagnosis antigen is characterized in that the nucleotide sequence is as shown in SEQ ID NO: 1. A primer pair for amplifying the nucleotide sequence is characterized in that an upstream primer sequence is as shown in SEQ ID NO: 3, and a downstream primer sequence is as shown in SEQ ID NO: 4. The invention further discloses application of the nucleotide sequence, the amino acid sequence and the BruAb20955 protein in preparation of a DIVA diagnostic reagent for brucellosis. The invention further discloses application of the nucleotide sequence, the amino acid sequence and the BruAb20955 protein in preparation of brucellosis prevention and control vaccines. According to the invention, a new molecular marker which can be used for differential diagnosis of Brucella wild virus infection and vaccine immunity is screened and identified at home and abroad for the first time, and meanwhile, an indirect ELISA method which can be used for experimental differential diagnosis is established by utilizing the new marker molecule. The judgment critical value of the method is 1.908.

The application provides a kind of anti-RSV N protein monoclonal antibody and its preparation method and application, belong to monoclonal antibody technical field.The heavy chain variable region amino acid sequence of the monoclonal antibody described in the application is as shown in SEQ ID NO.2, the light chain variable region amino acid sequence is as shown in SEQ ID NO.4, is secreted by hybridoma cell strain 1A2, subtype is IgG1, Kappa type, has high affinity and strong specificity to RSV N protein.The antibody can specifically recognize RSV N protein and inactivated RSV A type virus, has high titer, good stability, and can be used for preparing respiratory syncytial virus detection reagent, especially suitable for double antibody sandwich ELISA detection kit, provides core material for early rapid detection of RSV infection, has important clinical application value.

The application discloses a kit for multiple nucleic acid detection based on CRISPR / Cas12a combined with universal crRNA, and belongs to the field of nucleic acid detection, and comprises crRNA, Cas12a, buffer 3.1, buffer 2, RNAase inhibitor, Bst DNA polymerase and dNTP mixed solution. The application utilizes the Cas12a trans-cleavage activity, and for the first time, a single type of crRNA is applied to nucleic acid detection of different types of viruses. Using the same crRNA, multiple viral nucleic acids, including but not limited to SARS-CoV-2, multiple subtypes of HPV viruses and tumor marker miRNA, etc. can be detected. The application widens the design ideas and methods of CRISPR / Cas nucleic acid detection, and has good analysis performance. Under optimized experimental conditions, the same crRNA can be used to effectively detect multiple DNA viruses and RNA viruses, and has high sensitivity, and can detect a concentration of 5 pM. Taking HPV-16 pseudovirus as an example, actual sample detection is carried out.

The application provides a kind of real-time monitoring system and method based on wild animal host influenza virus, it is related to animal influenza virus monitoring technical field, comprising the following steps: step S1, dynamic monitoring point is laid out;Step S2, sample collection and pretreatment;Step S3, real-time detection and typing virus;Step S4, data real-time transmission and sharing;Step S5, virus sequence alignment and variation analysis;Step S6, dynamic risk assessment and early warning.The real-time monitoring system and method based on wild animal host influenza virus, through portable real-time fluorescent RT-PCR detection equipment realizes field real-time detection, combines double mode communication and cloud real-time analysis, eliminates the time lag of sample transportation and laboratory detection, realizes the real-time feedback of virus information.

The invention provides an in-vivo and in-vitro double visual recombinant mouse poxvirus genome, a recombinant mouse poxvirus strain and application, and belongs to the technical field of gene engineering. The invention provides a recombinant mouse poxvirus genome. The recombinant mouse poxvirus genome comprises a mouse poxvirus genome, a red fluorescent protein gene mKate and a Gaussian luciferase protein gene Gluc, wherein the red fluorescent protein gene mKate and the Gaussian luciferase protein gene Gluc are inserted between the mouse poxvirus genome A11R and the mouse poxvirus genome A12L. According to the invention, the area A of the mouse poxvirus is modified, and the double-reporter gene is inserted between A11R and A12L for the first time, so that the mouse poxvirus-resistant high-throughput drug screening method can be used for anti-orthopoxvirus high-throughput drug screening, the in-vivo effect of the drug can be more intuitively displayed, and the operation is simple and convenient. The recombinant virus obtained by the invention has good hereditary stability, has similar biological characteristics and virulence to a wild type virus, can visualize infection and distribution of the virus in a mouse body, and can be used for in-vitro high-throughput screening of drugs and development of corresponding vaccines.

The invention is applicable to the technical field of genetic engineering, and provides an E2-1d-mi3 fusion protein of bovine viral diarrhea virus and a preparation method and application thereof, the E2-1d-mi3 fusion protein is formed by fusing a mi3 tag sequence with a 1d type E2 genome of the bovine viral diarrhea virus; the amino acid sequence of the mi3 tag sequence is as shown in a sequence table SEQ ID NO: 4; the amino acid sequence of the E2-1d-mi3 fusion protein is as shown in SEQ ID NO: 2 in a sequence table. After animals are immunized by the E2-1d-mi3 fusion protein, an effective immune protection effect can be generated, and the E2-1d-mi3 fusion protein has a good prevention and control application prospect on bovine viral diarrhea caused by BVDV 1d type viruses.

This invention discloses a Newcastle disease virus (NDV) genotype VII attenuated marker vaccine strain, rMG7-HN-G256E, with accession number CGMCC No. 46292. The complete nucleotide sequence of this vaccine strain is shown in SEQ ID No: 1; wherein, the 256th amino acid (G) of the HN protein is replaced with glutamic acid (E). This invention utilizes a Newcastle disease virus reverse genetics platform to mutate different sites on the MG7 Newcastle disease virus strain, rescuing a genotype VII Newcastle disease virus marker vaccine strain, rMG7-HN-G256E, which exhibits low pathogenicity, good growth characteristics, high viral titer, stable passage, and excellent immunogenicity. This marker vaccine strain has good immunogenicity, can induce high levels of protective antibodies, and provides complete protection for immunized chickens. It can be used to control the currently prevalent genotype VII Newcastle disease virus and lays the foundation for differentiating between vaccine immunization and wild-type virus infection.