# US 8,202,977 B2 · Pestivirus species > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,202,977 B2 - Document ID: US8202977B2 - Status: Granted - Kind: B2 - Application number: 12/298,026 - Filed: 2007-04-20 - Published: 2010-08-12 - Granted: 2012-06-19 - Earliest claimed: 2006-04-21 - Assignees: Minister for Primary Industries For And On Behalf Of The State Of New South Wales - Inventors: Melinda Jane Frost; Peter Daniel Kirkland; Deborah Susan Finlaison - Web page: https://patentgrep.com/patent/US8202977B2 ## CPC - C12Q 1/701: {Specific hybridization probes} - A61K 39/12: Viral antigens - A61P 31/12: Antivirals - A61P 31/14: for RNA viruses - C07K 14/005: from viruses - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - G01N 33/56983: {Viruses} - A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) - A61K 2039/53: {DNA (RNA) vaccination} - C12N 2770/24321: Viruses as such, e.g. new isolates, mutants or their genomic sequences - C12N 2770/24322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2770/24334: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12Q 2600/136: Screening for pharmacological compounds - G01N 2333/18: Togaviridae; Flaviviridae - G01N 2469/20: Detection of antibodies in sample from host which are directed against antigens from microorganisms - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The application relates to a pestivirus, designated PMC virus, that is associated with porcine myocarditis syndrome, and the gene and protein sequences derived therefrom. The application further relates to detection methods, vaccine therapeutics, and diagnostic methods using the PMC virus or gene/protein sequences derived therefrom. ## Claims ### Claim 1 An isolated RNA or DNA nucleotide sequence comprising: a) SEQ ID NO:11, wherein when the isolated RNA or DNA nucleotide sequence is an RNA nucleotide sequence, thymidine (t) nucleotides are substituted with uridine (u) nucleotides; b) an RNA or DNA sequence having at least 85% sequence identity with SEQ ID NO: 11; or c) an RNA or DNA sequence comprising the complement of the nucleotide sequence of (a) or (b). ### Claim 2 (depends on 1) A method for detecting the presence or absence of porcine myocarditis syndrome (PMC) virus in a biological sample, comprising the steps of: a) bringing the biological sample into contact with a polynucleotide probe or primer comprising an isolated RNA or DNA nucleotide sequence according to claim 1 under suitable hybridizing conditions; and b) detecting any duplex formed between the probe or primer and nucleic acid sequences in the sample. ### Claim 3 (depends on 1) A method for the detection of PMC virus nucleic acids present in a biological sample, comprising: a) amplifying the RNA or DNA nucleotide sequence of claim 1 with at least one primer, and b) detecting the amplified nucleic acids. ### Claim 4 (depends on 1) A method for the detection of PMC virus nucleic acids present in a biological sample, comprising: a) hybridizing the nucleic acids of the biological sample at appropriate conditions with one or more probes comprising an isolated RNA or DNA nucleotide sequence according to claim 1, b) washing under appropriate conditions, and c) detecting the hybrids formed. ### Claim 5 (depends on 1) A method for detecting viral RNA or DNA comprising the steps of: a) immobilizing PMC virus on a support; b) disrupting the virion; and c) hybridizing the nucleic acids of the virion with a probe comprising an isolated RNA or DNA nucleotide sequence according to claim 1. ### Claim 6 (depends on 1) A method for screening the tissue of subjects for PMC virus comprising the steps of: a) extracting DNA from tissue; b) restriction enzyme cleavage of said DNA; c) electrophoresis of the fragments; and d) Southern blotting of genomic DNA from tissues and subsequent hybridization with a labelled cloned PMC virus DNA sequence according to claim 1. ### Claim 7 (depends on 1) An immunogenic composition comprising an isolated RNA or DNA nucleotide sequence according to claim 1. ### Claim 8 (depends on 1) A vector comprising: (a) an isolated polynucleotide sequence encoding a PMC virus comprising an isolated RNA or DNA nucleotide sequence according to claim 1; and (b) a heterologous polynucleotide. ### Claim 9 (depends on 8) The vector of claim 8 wherein the heterologous polynucleotide is operably linked to the polynucleotide sequence of the PMC virus, such that expression of the polynucleotide sequence of the PMC virus also leads to expression of the heterologous polynucleotide sequence. ### Claim 10 (depends on 8) The vector of claim 8, wherein the PMC virus expressed from said vector is attenuated. ### Claim 11 (depends on 1) A kit for demonstrating the presence of PMC virus comprising: (a) a predetermined amount of at least one labelled nucleic acid sequence derived from the PMC virus, wherein said nucleic acid sequence comprises an isolated RNA or DNA nucleotide sequence according to claim 1; and (b) directions for use of said kit. ### Claim 12 (depends on 1) A recombinant expression vector comprising an isolated RNA or DNA nucleotide sequence of claim 1, operably linked to prokaryotic, eukaryotic or viral transcription and translation control elements. ### Claim 13 (depends on 12) An isolated host cell transformed by the vector of claim 12. ### Claim 14 (depends on 1, 12) A method for preparing a PMC virus amino acid sequence encoded by an isolated RNA or DNA nucleotide sequence according to claim 1, comprising the steps of: (a) culturing a host cell containing a vector according to claim 12 under conditions that provide for expression of the PMC virus amino acid sequence; and (b) recovering the expressed PMC virus sequence. ### Claim 15 (depends on 7) The composition of claim 7 wherein the composition further comprises a pharmaceutically acceptable carrier or diluent and/or an adjuvant. ### Claim 16 (depends on 7) A method of inducing an immune response in an animal comprising the steps of: providing an immunogenic composition according to claim 7 to the animal in an amount effective for producing an immune response. ### Claim 17 (depends on 16) The method of claim 16 wherein the immunogenic composition is administered by a method selected from the group consisting of: intravenously, subcutaneously, intramuscularly, intraorbitally, ophthalmically, intraventricularly, intracranially, intracapsularly, intraspinally, intracisternally, intraperitoneally, buccal, rectally, vaginally, intranasally, orally and aerosol administration. ### Claim 18 (depends on 12) A method for preparing a PMC virus amino acid sequence, comprising the steps of: (a) culturing a host cell containing a vector according to claim 12 under conditions that provide for expression of the PMC virus amino acid sequence; and (b) recovering the expressed PMC virus sequence.