Metapneumovirus strains and their use in vaccine formulations and as vectors for expression of antigenic sequences
Provided is an isolated mammalian negative strand RNA virus, metapneumovirus (MPV), within the sub-family Pneumoviridae, of the family Paramyxoviridae. Also provided are isolated mammalian negative strand RNA viruses identifiable as phylogenetically corresponding or relating to the genus Metapneumovirus and components thereof. In particular, provided is a mammalian MPV, subgroups and variants thereof. Also provided are genomic nucleotide sequences of different isolates of mammalian MPV, in particular, human MPV. Disclosed is the use of the sequence information of different isolates of mammalian MPV for diagnostic and therapeutic methods. Provided are nucleotide sequences encoding the genome of an MPV or a portion thereof, including both mammalian and avian MPV. Further described are chimeric or recombinant viruses encoded by the nucleotide sequences and chimeric and recombinant mammalian MPV that comprise one or more non-native or heterologous sequences. Also provided are vaccine formulations comprising mammalian or avian MPV, including recombinant and chimeric forms thereof. The vaccine preparations encompass multivalent vaccines, including bivalent and trivalent vaccine preparations.
1. 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 DNA probe does not specifically hybridize to a polynucleotide from avian pneumovirus (APV);
wherein the probe does not hybridize under stringent conditions to a sequence encoding a polypeptide of SEQ ID NOs:378 and 380 or the complement of the sequence;
wherein the stringent conditions include a hybridization buffer comprising 6×SSC and 1 mM EDTA; and
wherein the target polynucleotide comprises a first nucleic acid encoding a polypeptide that is at least 90% identical to SEQ ID NO:379 or the complement of the first nucleic acid; or
wherein the target polynucleotide comprises a second nucleic acid encoding a polypeptide that is at least 90% identical to SEQ ID NO:381 or the complement of the second nucleic acid.
2. The kit of claim 1 , wherein the DNA probe comprises at least 25 nucleotides.
3. The kit of claim 1 , wherein the DNA probe comprises at least 40 nucleotides.
4. The kit of claim 1 , wherein the DNA probe further comprises a detectable marker.
5. The kit of claim 1 , wherein the target polynucleotide comprises a nucleic acid encoding SEQ ID NO:375 or wherein the target polynucleotide comprises a nucleic acid encoding SEQ ID NO:377.
6. A method for detecting metapneumovirus (MPV) subtype A2 or B2 in a mammalian subject, the method comprising:
contacting a sample from the subject with a probe nucleic acid of at least 10 nucleotides that hybridizes under stringent conditions to a sequence encoding a polypeptide that is at least 90% identical to one of SEQ ID NOs:379 and 381 or the complement of the sequence; and
detecting hybridization of the probe and the target polynucleotide;
wherein the probe does not hybridize under stringent conditions to a sequence encoding a polypeptide of SEQ ID NOs: 378 and 380 or the complement of the sequence; and
wherein the stringent conditions include a hybridization buffer comprising 6×SSC and 1 mM EDTA.
7. The method according to claim 6 , wherein the probe nucleic acid comprises at least 25 nucleotides.
8. The method according to claim 6 , wherein the probe nucleic acid comprises at least 40 nucleotides.
9. The method according to claim 6 , wherein detecting hybridization of the probe and the target polynucleotide comprises performing PCR with the probe nucleic acid as a primer.
10. The method according to claim 6 , wherein the mammalian subject is a human.
11. The method according to claim 6 , wherein the probe nucleic acid comprises a detectable marker.
12. The method according to claim 6 , wherein the probe is attached to a solid support.
13. The method according to claim 6 , wherein the target polynucleotide comprises a nucleic acid encoding one of SEQ ID NOs: 375 and 377.
14. The method according to claim 6 , wherein the target polynucleotide comprises the complement of a nucleic acid encoding one of SEQ ID NOs: 375 and 377.
15. The method according to claim 6 , the method further comprising:
diagnosing the subject as infected with MPV based on the hybridization of the probe to the target polynucleotide.