Determination of hepatitis C virus genotype
View Patent ↗The present invention provides compositions and methods for the detection and characterization of HCV 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, to determine HCV genotype.
1. A composition comprising an invasive cleavage detection assay, wherein said invasive cleavage detection assay comprises two or more probe sets, wherein each probe set comprises a first and second oligonucleotide configured to form an invasive cleavage structure to detect at least a single nucleotide polymorphism in a position of a 5′ UTR sequence of HCV and wherein said first oligonucleotide comprises a 5′ portion and a 3′ portion, wherein said 3′ portion is configured to hybridize to said 5′ UTR sequence of HCV, and wherein said 5′ portion is configured to not hybridize to said 5′ UTR sequence of HCV, wherein said first oligonucleotide is a sequence selected from the group consisting of SEQ ID NOs: 2, 33, 34, 36, and 63, wherein nucleotide positions in said 5′ UTR are numbered in accordance with the consensus sequences SEQ ID NOS:64, 65, 66, 67, 68 and 69, and wherein said position of said at least a single nucleotide polymorphism is selected from the group consisting of: −245, −167, −163, −155, −144, −118, and −80, wherein position −245 is at nucleotide 30 in SEQ ID NOS:64-69; wherein position −167 is at nucleotide 108 of SEQ ID NOS:64-69; wherein position −163 is at nucleotide 112 of SEQ ID NOS:64-69; wherein position −155 is at nucleotide 120 of SEQ ID NOS:64-69; wherein position −144 is at nucleotide 131 in SEQ ID NO:69 and is not present in SEQ ID NOS:64-68; wherein position −118 is at nucleotide 157 in SEQ ID NOS:64-68 and nucleotide 160 in SEQ ID NO:69; and wherein position −80 is at nucleotide 195 in SEQ ID NOS:64-68 and nucleotide 198 in SEQ ID NO:69.
2. A composition comprising an invasive cleavage detection assay, wherein said invasive cleavage detection assay comprises two or more probe sets, wherein each probe set comprises a first and second oligonucleotide configured to form an invasive cleavage structure to detect at least a single nucleotide polymorphism in a position of a 5′ UTR sequence of HCV, wherein said second oligonucleotide comprises a 5′ portion and a 3′ portion, wherein said 5′ portion is configured to hybridize to said 5′ UTR sequence of HCV, and wherein said 3′ portion is configured to not hybridize to said 5′ UTR sequence of HCV and wherein said second oligonucleotide is a sequence selected from the group consisting of SEQ ID NOs:1, 31, 32, 35, and 56, wherein nucleotide positions in said 5′ UTR are numbered in accordance with the consensus sequences SEQ ID NOS:64, 65, 66, 67, 68 and 69 and wherein said position of said at least a single nucleotide polymorphism is selected from the group consisting of: −245, −167, −163, −155, −144, −118, and −80, wherein position −245 is at nucleotide 30 in SEQ ID NOS:64-69; wherein position −167 is at nucleotide 108 of SEQ ID NOS:64-69; wherein position −163 is at nucleotide 112 of SEQ ID NOS:64-69; wherein position −155 is at nucleotide 120 of SEQ ID NOS:64-69; wherein position −144 is at nucleotide 131 in SEQ ID NO:69 and is not present in SEQ ID NOS:64-68; wherein position −118 is at nucleotide 157 in SEQ ID NOS:64-68 and nucleotide 160 in SEQ ID NO:69; and wherein position −80 is at nucleotide 195 in SEQ ID NOS:64-68 and nucleotide 198 in SEQ ID NO:69.
3. The composition of claim 1 , wherein said 5′ UTR sequence of HCV comprises RNA.
4. The composition of claim 1 , wherein said 5′ UTR sequence of HCV comprises DNA.
5. The composition of claim 2 , wherein said 5′ UTR sequence of HCV comprises RNA.
6. The composition of claim 2 , wherein said 5′ UTR sequence of HCV comprises DNA.