40 results about "Influenza virus RNA" patented technology

The invention provides a method for detecting fowl influenza virus and parting primer system and augmenting fowl influenza virus RNA asymmetrically. The invention designs and screens 25 pairs multiple asymmetrical RT-PCR primers, a pair general primer, 52 specific probes and 3 quality control probes according to the report fowl influenza virus total hypotype genome sequence in Genebank, wherein every pair in 25 pairs multiple asymmetrical RT-PCR primers is fit for a hypotype of fowl influenza virus, the primer system comprising 25 pairs multiple asymmetrical RT-PCR primers and a pair general primer can be used for detecting and parting fowl influenza virus, the primer system builds the method of asymmetrically augmenting fowl influenza virus RNA and detecting and parting fowl influenza virus. The invention provides strong specificity, high sensibility, which also proceeds the first step application.

The present invention aims to express influenza virus RNA polymerase on a large scale, to crystallize the influenza virus RNA polymerase, and to provide a method for screening a substance capable of serving as an active ingredient in anti-influenza drugs which target a protein highly conserved among influenza virus species.The present invention provides a complex comprising a polypeptide consisting of an amino acid sequence at positions 239-716 of the RNA polymerase PA subunit in influenza A / Puerto Rico / 8 / 1934 H1N1 and a polypeptide consisting of an amino acid sequence at positions 1-81 of the RNA polymerase PB1 subunit in influenza A / Puerto Rico / 8 / 1934 H1N1, as well as a method for screening a substance capable of serving as an active ingredient in anti-influenza drugs, which comprises the step of selecting a substance which inhibits the interaction between α-subunit and β-subunit 1, each constituting influenza virus RNA polymerase, in the presence of a candidate substance.

The invention provides a combined nucleic acid real-time fluorescent detection method for simultaneously detecting an influenza A H1N1 virus and an influenza A virus and a kit. The combined nucleic acid real-time fluorescent detection method for the influenza A H1N1 virus and the influenza A virus comprises the following steps of: (1) extracting virus RNA; (2) carrying out fluorescent quantitative PCR (Polymerase Chain Reaction) detection; and (3) judging a detecting result. By carrying out multiple sequence comparison and aiming at a conserved gene fragment of the influenza A virus and the influenza A H1N1 virus (infecting in 2009), a primer and a probe with high specificity are designed and used for real-time fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection. The invention can be used for detecting influenza A virus RNA of human influenza, swine influenza, avian influenza, and other influenza viruses, meanwhile specifically detecting the influenza A H1N1 virus (infecting in 2009) RNA and carrying out double analysis so that a detecting result is more reliable.

The invention belongs to the technical field of gene engineering, and particularly relates to an A549-5Ps cell line for stable coexpression of influenza virus A RNA polymerases PB2, PB1, PA subunit and NP protein, and resistance reporter genes of Gaussia luciferase / blasticidin, and a building method and an application of the A549-5Ps cell line. The cell line has the characteristic of monitoring replication and expression of influenza virus A genome RNA in cells in real time; and a simple and sensitive method is provided for further revelation of the transcription and replication mechanism of the influenza virus A in the cells, illumination of the interaction between viruses and hosts, and research and development of anti-influenza virus A medicines.

The invention provides a one-step fluorescent parting RT-PCR detection kit for sendai virus. The kit comprises an RT-PCR reaction liquid, an RT-PCR enzyme mixture, a sendai virus parting primer probe,an internal reference, negative control, critical positive control and strong positive control. The kit can be used for carrying out one-step RT-PCR reaction directly on extracted sendai virus RNA and fluorescent parting detection on the sendai virus RNA in a sample, and preventing pollution by taking an internal reference gene sequence as internal control by means of a UNG enzyme. The one-step amplification method of the kit is simple, short in progress, simple to operate and pollution-preventative, and the detection result is high in specificity, high in sensitivity, clear in result and high in credibility. The kit can be used for fluorescent parting detection of type 1, type 2 and type 3 sendai viruses in a human nasopharyngeal swab sample.

The invention discloses a novel application of a caffeoylquinic acid compound. The novel application of caffeoylquinic acid compound provided in the invention shown in a formula I is an application of being as an influenza virus RNA polymerase inhibitor on one hand; on the other hand, the novel application of caffeoylquinic acid compound is an application of preparing the medicament for preventing and / or treating the diseases relative to the influenza virus RNA polymerase, especially relates to an application of preparing the medicament in resisting influenza virus, more especially relates toan application of preparing the medicament in resisting bird flu virus (H5N1).

The invention relates to generic methods for the detection and quantification of influenza viruses. These may uses a reverse transcription (RT-PCR) real time (q-PCR) assay which amplifies a conserved region within influenza A or B strains. The assays allow the quantification of influenza virus RNA molecules or whole virus particles, irrespective of the particular virus strain (e.g. human, avian, swine flu). The methods are particularly applicable as diagnostic assays or in the monitoring of vaccine production processes.

The invention relates to a kit for detecting product DNA of AH1N1 influenza virus DNA amplified through a loop-mediated isothermal nucleic acid amplification technology by using gold nanoparticles, belonging to the technical field of biological detection. In combination with the loop-mediated isothermal nucleic acid amplification technology and a biological nanotechnology, the invention has the advantages that the sensitivity of biological molecular detection is improved, the operation of the method is simple, the detection is rapid, the accuracy is high, special instruments and devices are not required, and the kit can be widely used for high-sensitivity AH1N1 influenza virus detection in fields such as family diagnosis, clinical diagnosis, infectious disease control, environmental monitoring, inspection and quarantine, biotechnology and the like. The kit comprises: (1) gold nanoparticles labeled with molecular probes capable of identifying specific sequences of AH1N1 influenza viruses; and (2) a loop-mediated isothermal nucleic acid amplification system capable of amplifying AH1N1 influenza virus DNA in a sample to be detected or capable of amplifying AH1N1 influenza virus RNA in the sample to be detected after reverse transcription.

Silencing of Influenza virus RNA can be achieved by siDNA. These are oligodeoxynucleotides having an antisense-strand homologous to the viral RNA and a second strand, partially complementary to the antisense-strand. The two strands are preferentially linked by a linker (eg 4 thymidines). Triple-helix formation with the target RNA is a preferred effect. The siDNA is superior to siRNA because it acts earlier, is easier taken up by the cell, the formation of RNA-DNA hybrids is preferred over double-stranded DNA or double-stranded RNA, which forms as tertiary structures in RNA genomes. Also the induction of interferon is less likely. siDNA is easier to synthesize and it is more stable. It can be combined with siRNA.

The invention belongs to the field of biological detection and discloses a kit for rapidly extracting RNA of influenza viruses. The kit comprises virus splitting and binding liquid which contains polyether alcohol, sucrose and Tris-HCl. The kit disclosed by the invention has the advantages that by utilization of the kit, the nucleic acid of the influenza viruses can be released only by one-step operation, and the following substances can be used for fluorescence PCR (Polymerase Chain Reaction) detection.

The invention discloses an influenza virus reverse transcription loop-mediated isothermal amplification (RT-LAMP) kit and application thereof. The kit comprises a RT-LAMP primer, 2* reaction buffer solution, enzyme mixture EM, a fluorescence visual detection reagent, ultrapure water and an influenza virus RNA template, wherein the RT-LAMP primer comprises outer primers F3 and B3, and inner primers FIP and BIP. Specificity detection, sensitivity detection and fluorescence visible detection prove that the RT-LAMP detection method can specifically detect influenza virus, can be used for monitoring reactions in real time and quantitatively detecting the number of influenza virus copies so as to quickly and correctly acquire the detection result, and brings convenience to simple, quick and reliable influenza virus detection.

The invention discloses a hydroxypyridinone [1, 2b] [1, 2, 5] triazepine derivative as well as preparation and application thereof. Experiments prove that the hydroxypyridinone [1, 2b] [1, 2, 5] triazepine derivative (as shown in the general formula I) has a relatively good inhibition effect on the activity of influenza A virus RNA polymerase, and can be used as an influenza virus RNA polymerase inhibitor for treating cold caused by influenza viruses. The structural general formula I is shown in the specification,.

The invention discloses a kit for detecting a canine parainfluenza virus. The kit comprises a light shielding detection box, ALL-READY PCR BEADS full premix freeze-dried powder, a positive control group, a negative control group and a photodetector, the light shielding detection box and the photodetector are arranged in a fitting manner, a photoelectric detection module is arranged in the photoelectric detector, and the ALL-READY PCR BEADS full premix freeze-dried powder comprises transcriptase, a nucleic acid amplifier enzyme, a reaction buffer solution, a specific primer and a probe which are needed by the fluorescent quantitative PCR detection of the canine parainfluenza virus RNA; and the negative control group adopts canine parainfluenza virus RNA-free enzyme water, the positive control group is in vitro transcribed parainfluenza virus RNA, and the photoelectric detection module includes a photodiode. The design is optimized, the light shielding detection box is combined, and the photoelectric detection module is optimized, so the detection error is reduced, and the detection specificity is improved.

The invention discloses an anti-influenza virus function of flavaspidic acid AB. The invention provides application of the flavaspidic acid AB in preparation of at least one of the following products1)-5): 1) an influenza virus inhibitor; 2) a product for treating influenza; 3) a product for inhibiting proliferation or replication of influenza viruses; 4) a product for inhibiting synthesis of influenza virus RNA; and 5) a product for inhibiting expression or synthesis of a gene in the influenza virus. Experiments prove that the flavaspidic acid AB is extracted from male fern rhizome, the flavaspidic acid AB is a monomer compound from plants, and the flavaspidic acid AB can be used for preparing the influenza virus inhibitor, so that the flavaspidic acid AB is used for preventing and treating infection of influenza, human health is protected, and a new choice is provided for development of an influenza virus drug.

The present invention aims to express influenza virus RNA polymerase on a large scale, to crystallize the influenza virus RNA polymerase, and to provide a method for screening a substance capable of serving as an active ingredient in anti-influenza drugs which target a protein highly conserved among influenza virus species.The present invention provides a complex comprising a polypeptide consisting of an amino acid sequence at positions 239-716 of the RNA polymerase PA subunit in influenza A / Puerto Rico / 8 / 1934 H1N1 and a polypeptide consisting of an amino acid sequence at positions 1-81 of the RNA polymerase PB1 subunit in influenza A / Puerto Rico / 8 / 1934 H1N1, as well as a method for screening a substance capable of serving as an active ingredient in anti-influenza drugs, which comprises the step of selecting a substance which inhibits the interaction between α-subunit and β-subunit 1, each constituting influenza virus RNA polymerase, in the presence of a candidate substance.

The present invention provides the application of salvianolic acid B in the preparation of medicines for preventing or treating H5N1 influenza virus, wherein the salvianolic acid B has the following structural formula: salvianolic acid B can effectively inhibit the RNA polymerase subunit of H5N1 influenza virus Activity of PAN protein.

The invention provides an antiviral pharmaceutical molecule capable of inhibiting the activity of influenza virus RNA polymerase. An antiviral drug contains the structure represented by the followingformula 1, or the antiviral drug contains a compound with the structure represented by the formula 1 as a drug precursor.

The invention discloses a reporter gene plasmid for testing RNA (Ribonucleic Acid) polymerase activity of influenza viruses in canine cells and application thereof. According to the plasmid disclosed by the invention, the nucleotide sequence is shown in SEQ ID No. 1. In a dual-reporting genetic testing method, by adopting the reporter gene plasmid pGLucCa constructed by the invention, the RNA polymerase activity of the influenza viruses in the canine cells can be accurately reflected, errors caused by lysed cells can be reduced, cell supernatant can be extracted at different time points so as to carry out RNA polymerase activity assay, and then, the dual-reporting genetic RNA polymerase activity testing method is simpler and more convenient and is wider in range of application.

The invention discloses a reporter gene plasmid for testing RNA (Ribonucleic Acid) polymerase activity of influenza viruses in avian cells and application thereof. According to the plasmid disclosed by the invention, the nucleotide sequence is shown in SEQ ID No. 1. In a dual-reporting genetic testing method, by adopting the reporter gene plasmid pGLucCk constructed by the invention, the RNA polymerase activity of the influenza viruses in the avian cells can be accurately reflected, errors caused by lysed cells can be reduced, cell supernatant can be extracted at different time points so as to carry out RNA polymerase activity assay, and then, the dual-reporting genetic RNA polymerase activity testing method is simpler and more convenient and is wider in range of application.

The invention relates to a preparation method of an avian influenza virus inhibition material for human. The preparation method has the advantages that the raw material sources are wide; the production cost is low; the large-range popularization and application can be realized. The avian influenza virus inhibition material for human is sprinkled in regions in which persons and vehicles come and go, so that the avian influenza virus inhibition material is fully contact with avian influenza viruses; the avian influenza virus inhibition material can interfere the avian influenza virus RNA; the avian influenza virus replication is inhibited, so that the avian influenza virus is inactivated; the avian influenza virus propagation is prevented.

The invention provides a combined nucleic acid real-time fluorescent detection method for simultaneously detecting an influenza A H1N1 virus and an influenza A virus and a kit. The combined nucleic acid real-time fluorescent detection method for the influenza A H1N1 virus and the influenza A virus comprises the following steps of: (1) extracting virus RNA; (2) carrying out fluorescent quantitative PCR (Polymerase Chain Reaction) detection; and (3) judging a detecting result. By carrying out multiple sequence comparison and aiming at a conserved gene fragment of the influenza A virus and the influenza A H1N1 virus (infecting in 2009), a primer and a probe with high specificity are designed and used for real-time fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection. The invention can be used for detecting influenza A virus RNA of human influenza, swine influenza, avian influenza, and other influenza viruses, meanwhile specifically detecting the influenza A H1N1virus (infecting in 2009) RNA and carrying out double analysis so that a detecting result is more reliable.

The invention provides a method for detecting fowl influenza virus and parting primer system and augmenting fowl influenza virus RNA asymmetrically. The invention designs and screens 25 pairs multiple asymmetrical RT-PCR primers, a pair general primer, 52 specific probes and 3 quality control probes according to the report fowl influenza virus total hypotype genome sequence in Genebank, wherein every pair in 25 pairs multiple asymmetrical RT-PCR primers is fit for a hypotype of fowl influenza virus, the primer system comprising 25 pairs multiple asymmetrical RT-PCR primers and a pair general primer can be used for detecting and parting fowl influenza virus, the primer system builds the method of asymmetrically augmenting fowl influenza virus RNA and detecting and parting fowl influenza virus. The invention provides strong specificity, high sensibility, which also proceeds the first step application.

The invention provides an azaindole derivative containing aza amino acid as well as preparation and application thereof. The compound has a good inhibition effect on influenza viruses in vitro, IC50 and EC50 in anti-influenza virus polymerase and anti-influenza virus replication reach the nanomole level, the toxicity of the compound is small, CC50 of most of the compound on MDCK cells and A549 cells is 30 muM or above, and the selectivity of the compound is good. The compound provided by the invention aims at the PB2 subunit of influenza virus RNA polymerase, can be used for preparing anti-influenza drugs, can solve the problem of drug resistance of the existing clinical anti-influenza drugs, and has a relatively high application value. The structural general formula (I) of the derivative is shown in the specification,.

The invention discloses a subtype detection method of influenza A virus, which comprises the following steps: Step 1, designing and synthesizing primers; Step 2, preparing carboxyl magnetic beads connected with magnetic beads and forward primers; Step 3, A Influenza virus RNA extraction; Step 4, RT‑PCR; Step 5, magnetic adsorption and washing; Step 6, detection. The influenza A virus subtype detection method of the present invention can perform high-throughput, rapid, sensitive and specific typing of the influenza A virus subtype, and has good application value.

The invention provides an antiviral pharmaceutical molecule capable of inhibiting the activity of influenza virus RNA polymerase. An antiviral drug contains the structure represented by the followingformula 1, or the antiviral drug contains a compound with the structure represented by the formula 1 as a drug precursor.

The invention belongs to the technical field of medicinal chemistry, and particularly relates to an antiviral drug Musellarin and analogue molecule taking influenza A virus RNA polymerase as a target,a preparation method and application. The chemical structural general formula of the molecule is shown as a formula i in the specification, in the formula i, R1 is selected from hydroxyl or alkoxy, R2 is selected from hydroxyl or alkoxy, R3 is selected from hydrogen atom, hydroxyl or alkoxy, and R4 is selected from hydroxyl or alkoxy. According to the antiviral drug Musellarin and analogue molecule provided by the invention, the influenza virus RNA polymerase is taken as a target, the transcription and replication processes of influenza virus are directly blocked by inhibiting the activity ofthe influenza virus RNA polymerase, and the molecule has efficient inhibition property and high selectivity by taking the influenza virus RNA polymerase as the target, so that the molecule has the potential of being developed into an anti-influenza virus drug molecule.

The invention discloses an influenza virus RNA polymerase purifying or crystallizing method. The influenza virus RNA polymerase crystallizing method comprises the following steps: co-expressing and purifying PB1 subunit encoding gene and PA subunit encoding gene of influenza virus RNA polymerase and encoding gene of PB2 subunit truncation in insect cells to obtain RNA polymerase, combining the RNA polymerase with RNA to form a complex, and crystallizing the complex to obtain influenza virus RNA polymerase crystals, wherein the PB2 subunit truncation is a protein formed by amino acid residues from N end to 126th position in a PB2 subunit amino acid sequence. Experiments prove that after different PB2 subunit truncations are used, the expression level of the influenza virus polymerase complex containing the 126 amino acid fragments of the amino end of PB2 is high, and 10mg of the protein can be purified through 1L of an insect cell culture solution, so the expression level of the influenza virus polymerase complex containing the 126 amino acid fragments of the amino end of PB2 is higher than that of other truncations; and the crystals obtained in the invention have the advantages of good purity, slight degradation, good crystallization repeatability, good quality, large size and strong diffraction.

The invention discloses a method for crystallizing influenza virus RNA polymase, which comprises the steps of carrying out coexpression on a PB1 subunit encoding gene, a PA subunit encoding gene and a PB2 truncated subunit encoding gene of influenza virus RNA polymase in an insect cell to obtain RNA polymase; and purifying and crystallizing to obtain a influenza virus RNA polymase crystal. An experiment proves that PB2 truncated protein segments which have different lengths and amino ends retained and other two subunit full-length proteins are subjected to coexpression purification and complex assembling, a small truncated complex can use different strains to obtain crystal at multiple truncating circumstances. The method lays a foundation for structure analysis of influenza virus RNA polymase and also has important meanings for anti-influenza virus drug screening and design and research and development process of broad spectrum vaccines and drugs.