Compositions and methods for detecting or quantifying parainfluenza virus
View Patent ↗Compositions, methods, kits, and uses are provided for detecting or quantifying an Human Parainfluenza virus 1 (HPIV-1), HPIV-2, HPIV-3, and/or HPIV-4 nucleic acid, e.g., using nucleic acid amplification and hybridization assays. In some embodiments, the compositions, methods, kits, and uses target the HN gene of HPIV-1, HPIV-2, and/or HPIV-3 and/or the NP gene of HPIV-4.
1. A composition or kit comprising at least first and second amplification oligomers, wherein:
the first amplification oligomer and second amplification oligomer are configured to amplify an HPIV-3 amplicon of at least about 50 nucleotides in length comprising at least one Human Parainfluenza Virus 3 (HPIV-3) position located in the range of positions 1350-1360 and/or 1380-1390, wherein the first oligomer comprises the sequence of SEQ ID NO: 28;
the composition or kit optionally further comprises a third oligomer configured to hybridize to the amplicon; and:
at least one oligomer comprises a non-nucleotide detectable label; at least one oligomer comprises a non-Watson Crick (NWC) position; or at least one oligomer comprises at least one 5-methylcytosine.
2. A method of detecting HPIV-3 in a sample, comprising:
contacting the sample with at least first and second amplification oligomers, thereby forming a composition according to claim 1 which further comprises the sample,
performing a nucleic acid amplification reaction in the composition which produces an HPIV-3 amplicon of at least about 50 nucleotides in length in the presence of an HPIV-3 nucleic acid, and
detecting the presence or absence of the at least one amplicon.
3. The composition or kit of claim 1 , wherein the second oligomer competes for hybridization to an HPIV-3 nucleic acid under stringent conditions with an oligomer having a sequence consisting of SEQ ID NO: 37 and the third oligomer competes for hybridization to an HPIV-3 nucleic acid under stringent conditions with an oligomer having a sequence consisting of SEQ ID NO: 31, 32 or 33.
4. The composition or kit of claim 1 , wherein the second oligomer comprises the sequence of SEQ ID NO: 37.
5. The composition or kit of claim 1 , wherein the third oligomer competes for hybridization to an HPIV-3 nucleic acid under stringent conditions with an oligomer having a sequence consisting of SEQ ID NO: 31, 32, or 33.
6. The composition or kit of claim 1 , wherein the third oligomer comprises the sequence of SEQ ID NO: 31.
7. The composition or kit of claim 1 , wherein the third oligomer comprises the sequence of SEQ ID NO: 32.
8. The composition or kit of claim 1 , wherein the third oligomer comprises the sequence of SEQ ID NO: 33.
9. The composition or kit of claim 1 , wherein all or all but one or two of the C residues in one, two, or three of the first, second, and third oligomers are 5-methylated.
10. The composition or kit of claim 1 , further comprising a first Human Parainfluenza Virus 1 (HPIV-1) amplification oligomer and a second HPIV-1 amplification oligomer, wherein:
the first HPIV-1 amplification oligomer and second HPIV-1 amplification oligomer are configured to amplify an HPIV-1 amplicon of at least about 50 nucleotides in length comprising at least one HPIV-1 position located within HPIV-1 positions 330-490 or 960-1100.
11. The composition or kit of claim 1 , further comprising a first Human Parainfluenza Virus 1 (HPIV-2) amplification oligomer and a second HPIV-2 amplification oligomer, wherein:
the first HPIV-2 amplification oligomer and second HPIV-2 amplification oligomer are configured to amplify an HPIV-2 amplicon of at least about 50 nucleotides in length comprising at least one HPIV-2 position located within HPIV-2 positions 1600-1700.
12. The composition or kit of claim 1 , further comprising a first Human Parainfluenza Virus 1 (HPIV-4) amplification oligomer and a second HPIV-4 amplification oligomer, wherein:
the first HPIV-4 amplification oligomer and second HPIV-4 amplification oligomer are configured to amplify an HPIV-4 amplicon of at least about 50 nucleotides in length comprising at least one HPIV-4 position located within HPIV-4 positions 620-740, 2130-2410, 2520-3040, or 10090-11980.
13. The composition or kit of claim 1 , wherein the third oligomer hybridizes to an HPIV nucleic acid at an antisense version of the site to which an oligomer having a sequence consisting of SEQ ID NO: 31, 32, or 33 hybridizes.