Detection of HPV
View Patent ↗The present invention provides compositions and methods for the detection and characterization of HPV sequences. More particularly, the present invention provides compositions, methods and kits for using invasive cleavage structure assays (e.g. the INVADER assay) to screen nucleic acid samples, e.g., from patients, for the presence of any one of a collection of HPV sequences. The present invention also provides compositions, methods and kits for screening sets of HPV sequences in a single reaction container.
1. A kit comprising oligonucleotide detection assay components for detecting HPV sequences, wherein said components consist of:
a) first, second and third pools of oligonucleotide sets, said sets consisting of first and second oligonucleotides, wherein the combination of first and second oligonucleotides form an invasive cleavage structure in combination with a target sequence comprising a HPV sequence, wherein said first pool comprises a first oligonucleotide of SEQ ID NO. 77 and a second oligonucleotide of SEQ ID NO. 78; said second pool comprises a first oligonucleotide of SEQ ID NO. 127 and a second oligonucleotide of SEQ ID NO. 129; and wherein said third pool comprises a first oligonucleotide of SEQ ID NO. 160 and a second oligonucleotide of SEQ ID NO. 161;
b) a FEN-1 endonuclease;
c) an internal control;
d) a plurality of fluorescence resonance energy transfer (FRET) cassettes; and
e) assay reagents comprising a buffer solution.
2. The kit of claim 1 , wherein said first pool of oligonucleotides further comprise SEQ ID NO. 79 and SEQ ID NO. 80.
3. The kit of claim 1 , wherein said second pool of oligonucleotides further comprise SEQ ID NO. 91, SEQ ID NO. 94, SEQ ID NO. 142, SEQ ID NO. 90, and SEQ ID NO. 95.
4. The kit of claim 1 , wherein said third pool of oligonucleotides further comprise SEQ ID NO. 110, SEQ ID NO. 163, SEQ ID NO. 173, SEQ ID NO. 177, SEQ ID NO. 180, SEQ ID NO. 111, SEQ ID NO. 112, SEQ ID NO. 113, SEQ ID NO. 164, SEQ ID NO. 165, SEQ ID NO. 166 and SEQ ID NO. 175.
5. The kit of claim 1 , wherein each of said first and second oligonucleotides comprise at least a one base pair mismatch with said target sequence.
6. The kit of claim 1 , wherein said first and second oligonucleotides of each pool form invasive cleavage structures under the same reaction conditions.
7. The kit of claim 1 , wherein oligonucleotides of said first pool participate in a detectable invasive cleavage structure in the presence of a target sequence comprising HPV type 51 sequence and/or HPV type 56 sequence and no other high risk HPV sequence.
8. The kit of claim 1 , wherein oligonucleotides of said second pool participate in a detectable invasive cleavage structure in the presence of a target sequence comprising HPV type 18 sequence, HPV type 39 sequence, HPV type 45 sequence, HPV type 59 sequence, HPV type 68 sequence and/or HPV type 70 sequence and no other high risk HPV sequence.
9. The kit of claim 1 , wherein oliognucleotides of said third pool participate in a detectable invasive cleavage structure in the presence of a target sequence comprising HPV type 16 sequence, HPV type 31 sequence, HPV type 33 sequence, HPV type 35 sequence, HPV type 52 sequence, HPV type 58 and/or HPV type 67 sequence and no other high risk HPV sequence.
10. The kit of claim 1 , wherein said oligonucleotide detection assay components detect two or more HPV types simultaneously in the presence or absence of other, non-detected HPV types.