Virus causing respiratory tract illness in susceptible mammals
The invention relates to the field of virology. The invention provides an isolated essentially mammalian negative-sense single stranded RNA virus (MPV) within the sub-family Pneumovirinae of the family Paramyxoviridae and identifiable as phylogenetically corresponding to the genus Metapneumovirus and components thereof.
1. A method for identifying a viral isolate from a mammalian subject as comprising a human metapneumovirus , the method comprising:
reacting the viral isolate or a component thereof with a DNA molecule of at least 10 nucleotides in length that is at least 90% homologous to a stretch of the DNA sequence of SEQ ID NO: 36, or the complement thereof;
wherein reaction of the DNA molecule with the viral isolate or component thereof indicates the presence of human metapneumovirus in the viral isolate from the mammalian subject.
2. The method according to claim 1 , wherein the DNA molecule that is at least 90% homologous is at least 18 nucleotides in length.
3. The method according to claim 2 , wherein the DNA molecule of at least 18 nucleotides in length is at least 90% homologous to a stretch of nucleotides in bases 27-1277 of DNA sequence of SEQ ID NO: 36, or the complement thereof.
4. The method according to claim 1 , wherein the DNA molecule that is at least 90% homologous is at least 25 nucleotides in length.
5. The method according to claim 4 , wherein the DNA molecule of at least 25 nucleotides in length is at least 90% homologous to a stretch of nucleotides in bases 27-1277 of DNA sequence of SEQ ID NO: 36, or the complement thereof.
6. The method according to claim 1 , wherein the DNA molecule of at least 10 nucleotides in length is at least 90% homologous to a stretch of nucleotides in bases 27-1277 of DNA sequence of SEQ ID NO: 36, or the complement thereof.
7. The method according to claim 1 , wherein the DNA molecule comprises SEQ ID NO: 122.
8. The method according to claim 1 , further comprising allowing the foimation of a complex comprising the DNA molecule of at least 10 nucleotides and the DNA sequence, or the complement thereof and detecting the complex.
9. A method for diagnosing a human metapneumovirus infection in a mammal, the method comprising:
determining in a sample of the mammal the presence of a component of human metapneumovirus by reacting the sample with a DNA molecule of at least 10 nucleotides in length that is at least 90% homologous to a stretch of the DNA sequence of SEQ ID NO: 36, or the complement thereof;
wherein reaction of the DNA molecule with the sample of the mammal indicates human metapneumovirus infection of the mammal.
10. The method according to claim 9 , wherein the DNA molecule that is at least 90% homologous is at least 18 nucleotides in length.
11. The method according to claim 10 , wherein the DNA molecule of at least 18 nucleotides in length is at least 90% homologous to a stretch of nucleotides in bases 27-1277 of DNA sequence of SEQ ID NO: 36, or the complement thereof.
12. The method according to claim 9 , wherein the DNA molecule that is at least 90% homologous is at least 25 nucleotides in length.
13. The method according to claim 12 , wherein the DNA molecule of at least 25 nucleotides in length is at least 90% homologous to a stretch of nucleotides in bases 27-1277 of DNA sequence of SEQ ID NO: 36, or the complement thereof.
14. The method according to claim 9 , wherein the DNA molecule of at least 10 nucleotides in length is at least 90% homologous to a stretch of nucleotides in bases 27-1277 of DNA sequence of SEQ ID NO: 36, or the complement thereof.
15. The method according to claim 9 , wherein the DNA molecule comprises SEQ ID NO: 122.
16. A method for diagnosing a human metapneumovirus infection of a mammal, the method comprising:
determining in a sample from the mammal the presence of human metapneumovirus or component thereof by reacting the sample with a DNA molecule specifically reactive with a stretch of at least 10 nucleotides of the DNA sequence of SEQ ID NO: 36, or the complement thereof;
wherein reaction of the DNA molecule with the sample of the mammal indicates human metapneumovirus infection of the mammal.
17. The method according to claim 16 , wherein the DNA molecule is specifically reactive with a stretch of at least 18 nucleotides of the DNA sequence of SEQ ID NO: 36.
18. The method according to claim 16 , wherein the DNA molecule is specifically reactive with a stretch of at least 25 nucleotides of the DNA sequence of SEQ ID NO: 36.
19. A method of detecting if a polynucleotide from human metapneumovirus is present in a mammalian subject, the method comprising:
performing an nucleic acid amplification reaction on a sample derived from the mammalian subject utilizing a set of primers for the amplification of at least a portion of the N gene of human metapneumovirus to create an amplified product, wherein the N gene, when transcribed and translated, produces a polypeptide at least 90% homologous to SEQ ID NO: 1;
contacting the amplified product with a DNA molecule that hybridizes with the amplified product;
hybridizing the DNA molecule to the amplified product; and
detecting the hybridized DNA molecule;
wherein the detection of the hybridized DNA molecule indicates the presence of human metapneumovirus in the mammalian subject.
20. The method according to claim 19 , wherein the DNA molecule is at least 10 bases in length.
21. The method according to claim 19 , wherein the DNA molecule is at least 40 bases in length.
22. The method according to claim 19 , wherein the DNA molecule comprises SEQ ID NO: 122.
23. The method according to claim 19 , wherein the set of primers comprises SEQ ID NOs: 113 and 114.
24. The method according to claim 19 , wherein the N gene comprises bases 27-1227 of SEQ ID NO:36.
25. The method according to claim 19 , wherein the nucleic acid amplification reaction is selected from the group consisting of PCR, RAP, and RT-PCR.
26. The method according to claim 19 , wherein the DNA molecule comprises a label.
27. The method according to claim 26 , wherein detecting the hybridized DNA molecule comprises detecting the label.
28. A method of detecting if a polynucleotide from human metapneumovirus is present in a mammalian subject, the method comprising:
performing an nucleic acid amplification reaction on a sample derived from the mammalian subject utilizing a set of primers for the amplification of at least a portion of the N gene of human metapneumovirus to create an amplified product, wherein the N gene, when transcribed and translated, produces a polypeptide at least 90% homologous to SEQ ID NO: 1;
contacting the amplified product with a DNA molecule of at least 25 nucleotides in length that specifically hybridizes with the amplified product;
hybridizing the DNA molecule to the amplified product; and
detecting the hybridized DNA molecule;
wherein the detection of the hybridized DNA molecule indicates the presence of human metapneumovirus in the mammalian subject.