Determination of hepatitis C virus genotype
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 method for determining the genotype of hepatitis C virus in a sample comprising;
a) providing;
i) a sample from a subject, wherein said sample is suspected of containing hepatitis C virus, and
ii) two or more probe sets, wherein each of probe sets is configured for detecting a single nucleotide polymorphism in a 5′ UTR sequence of HCV, and
b) contacting said sample with said two or more probe sets under conditions such that at least partial genotype information for each of said single nucleotide polymorphisms is determined thereby generating a 5′ UTR genotype profile, wherein said 5′ UTR genotype profile comprises at least partial genotype information for single nucleotide polymorphisms located in at least two of the following positions: −245, −167, −163, −155, −144/5, −118, and −80; wherein said positions are found in SEQ ID NOs 64, 65, 66, 67, 68 and 69 as shown in FIG. 10 , and wherein said 5′ UTR genotype profile is sufficient to genotype said hepatitis C virus.
2. The method of claim 1 , further comprising c) genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype I, II, IV, V, or VI.
3. The method of claim 2 , wherein said genotyping classifies said hepatitis C virus as being Genotype I, IV or V.
4. The method of claim 1 , wherein said two or more probe sets comprise invasive cleavage structure type probes.
5. The method of claim 1 , wherein each probe set comprises a first oligonucleotide, 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.
6. The method of claim 1 , wherein each probe set comprises a second oligonucleotide, 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.
7. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) an A at position −163, and ii) no C at position −144, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype I.
8. The method of claim 7 , wherein said 5′ UTR genotype profile further comprises: iii) no C at position −167, or a T at position −167.
9. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) an A at position −163, and ii) a T at position −80, wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype I.
10. The method of claim 9 , wherein said 5′ UTR genotype profile further comprises: iii) no C at position −167, or a T a position −167.
11. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) an A at position −163, and ii) a C at position −72, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype I.
12. The method of claim 11 , wherein said 5′ UTR genotype profile further comprises: iii) no C at position −167, or a T a position −167.
13. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −122, and ii) no C at position −144, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype I.
14. The method of claim 13 , wherein said 5′ UTR genotype profile further comprises: iii) no C at position −167.
15. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −122, and ii) a T at position −80, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype I.
16. The method of claim 15 , wherein said 5′ UTR genotype profile further comprises: iii) no C at position −167.
17. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −122, and ii) a C at position −72, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype I.
18. The method of claim 17 , wherein said 5′ UTR genotype profile further comprises: iii) no C at position −167.
19. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) an A at position −132, and ii) a G at position −163, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype II.
20. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −245, and ii) a C at position −167, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype IV.
21. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −245, and ii) an A at position −118, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype IV.
22. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −167, ii) a G at position −163, and iii) a C at position −155, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype V.
23. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a C at position −122, and ii) a G at position −117, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype V.
24. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a G at position −117, and ii) a C at position −80, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype VI.
25. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) an A at position −118, and ii) a C at position −80, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype VI.
26. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −122, and ii) a C at position −80, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype VI.
27. The method of claim 1 , wherein said 5′ UTR genotype profile comprises: i) a T at position −122, and ii) a T at position −72, and wherein the method further comprises genotyping said hepatitis C virus based on said 5′ UTR genotype profile as Genotype VI.