Metapneumovirus strains and their use in vaccine formulations and as vectors for expression of antigenic sequences
The present invention provides an isolated mammalian negative strand RNA virus, metapneumovirus (MPV), within the sub-family Pneumoviridae, of the family Paramyxoviridae. The invention also provides isolated mammalian negative strand RNA viruses identifiable as phylogenetically corresponding or relating to the genus Metapneumovirus and components thereof. In particular the invention provides a mammalian MPV, subgroups and variants thereof. The invention relates to genomic nucleotide sequences of different isolates of mammalian metapneumoviruses, in particular human metapneumoviruses. The invention relates to the use of the sequence information of different isolates of mammalian metapneumoviruses for diagnostic and therapeutic methods. The present invention relates to nucleotide sequences encoding the genome of a metapneumovirus or a portion thereof, including both mammalian and avian metapneumovirus . The invention further encompasses chimeric or recombinant viruses encoded by said nucleotide sequences. The invention also relates to chimeric and recombinant mammalian MPV that comprise one or more non-native or heterologous sequences. The invention further relates to vaccine formulations comprising mammalian or avian metapneumovirus , including recombinant and chimeric forms of said viruses. The vaccine preparations of the invention encompass multivalent vaccines, including bivalent and trivalent vaccine preparations.
1. A method for determining the presence of metapneumovirus (MPV) in a mammalian subject, the method comprising:
contacting a sample from the subject comprising the target polynucleotide with a DNA probe of at least 10 nucleotides that hybridizes specifically to target polynucleotide;
wherein the target polynucleotide comprises a sequence encoding a polypeptide that is at least 90% identical to SEQ ID NO:366 or the complement of the sequence;
hybridizing the DNA probe and the target polynucleotide to form a complex; and
detecting the complex.
2. The method according to claim 1 , wherein the DNA probe does not specifically hybridize to a polynucleotide from avian pneumovirus (APV).
3. The method according to claim 1 , wherein the DNA probe does specifically hybridize to a polynucleotide from avian pneumovirus (APV).
4. The method according to claim 1 , wherein the DNA probe comprises at least 25 nucleotides.
5. The method according to claim 1 , wherein the DNA probe comprises at least 40 nucleotides.
6. The method according to claim 1 , wherein detecting the complex comprises performing PCR with the DNA probe as a primer.
7. The method according to claim 1 , wherein the mammalian subject is a human.
8. The method according to claim 1 , wherein the DNA probe comprises a detectable marker.
9. The method according to claim 1 , wherein the DNA probe is attached to a solid support.
10. The method according to claim 1 , wherein the target polynucleotide comprises a nucleic acid encoding SEQ ID NO:366.
11. The method according to claim 1 , wherein the target polynucleotide comprises the complement of a nucleic acid encoding SEQ ID NO:366.
12. A kit for determining the presence of metapneumovirus (MPV) in a mammalian subject, the kit comprising:
a DNA probe of at least 10 nucleotides that specifically hybridizes to a target polynucleotide,
wherein the target polynucleotide comprises a sequence encoding a polypeptide that is at least 90% identical to SEQ ID NO:366 or the complement of the sequence, and
wherein the DNA probe does not specifically hybridize to a polynucleotide from avian pneumovirus (APV).
13. The kit of claim 12 , wherein the DNA probe comprises at least 25 nucleotides.
14. The kit of claim 12 , wherein the DNA probe comprises at least 40 nucleotides.
15. The kit of claim 12 , wherein the DNA probe further comprises detectable marker.
16. The kit of claim 12 , wherein the target polynucleotide comprises a nucleic acid encoding SEQ ID NO:366.
17. The method according to claim 12 , wherein the target polynucleotide comprises the complement of a nucleic acid encoding SEQ ID NO:366.