Hybridization assay detection probes for detecting human papilloma virus in a sample
View Patent ↗Hybridization assay detection probes targeted to HPV Type 16 nucleic acid sequences which are particularly useful to aid in detecting HPV type 16 are described. The oligonucleotides can aid in detecting HPV Type 16 by acting singly or as part of a detection probe mixture. Compositions, reaction mixtures and methods of use are provided.
1. A method of detecting a HPV Type 16 nucleic acid in a sample comprising the steps of:
a) contacting an HPV type 16 amplification product with at least one hybridization assay detection probe that forms a detectable probe: HPV type 16 target hybrid under stringent hybridization conditions, wherein the at least one hybridization assay detection probe comprises a detectable label joined to a contiguous nucleic acid sequence that is from 21 to 27 nucleotides in length, (a) contained within a sequence consisting of SEQ ID NO:13 or an RNA equivalent thereof, and contains a sequence consisting of SEQ ID NO:34 or an RNA equivalent thereof, or (b) contained within a sequence consisting of SEQ ID NO:14 or an RNA equivalent thereof, and contains a sequence consisting of SEQ ID NO:33 or an RNA equivalent thereof; and
b) detecting a signal from the probe: HPV type 16 target hybrid, thereby indicating the presence of HPV type 16 in the sample.
2. The method of claim 1 , wherein the contiguous nucleic acid sequence comprises a sequence selected from the group consisting of: SEQ ID NO:13; SEQ ID NO:14; SEQ ID NO:15; and SEQ ID NO:16.
3. The method of claim 1 , wherein the contiguous nucleic acid sequence comprises a sequence selected from the group consisting of: SEQ ID NO:33; SEQ ID NO:34; SEQ ID NO:35; and SEQ ID NO:36.
4. The method of claim 1 , wherein at least one of the nucleotide residues in the contiguous nucleic acid sequence of the probe is a 2′Omethyl ribose.
5. The method of claim 1 , wherein a fluorescent detectable label is joined to the contiguous nucleic acid sequence.
6. The method of claim 1 , wherein a chemiluminescent detectable label is joined to the contiguous nucleic acid sequence.
7. The method of claim 6 , further comprising a helper probe.
8. The method of claim 1 , further comprising a helper probe.
9. The method of claim 1 , wherein the hybridization assay detection probe comprises at least one chemiluminescent detectable label joined to the contiguous nucleic acid sequence wherein at least one of the residues of the contiguous nucleic acid sequence is a 2′Omethyl ribose, and wherein the contiguous nucleic acid sequence is selected from the group consisting of: (i) a nucleic acid sequence that is from 21 to 27 nucleotides in length, is contained within a sequence consisting of an RNA equivalent of SEQ ID NO:13 and contains an RNA equivalent of SEQ ID NO:34; and (ii) a nucleic acid sequence that is 21 to 27 nucleotides in length, is contained within a sequence consisting of and RNA equivalent of SEQ ID NO:14 and contains an RNA equivalent of SEQ ID NO:33.
10. The method of claim 1 , wherein the sample is a clinical sample.
11. The method of claim 1 , wherein the hybridization assay detection probe forms a detectable probe: HPV type 16 target hybrid under selective stringency hybridization conditions.
12. The method of claim 1 , wherein the sample type is selected from the group consisting of: urine samples, tissue sections, urogenital secretions, and urogenital swabs.