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 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 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 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 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 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 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 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.