Hepatitis-C virus testing
AssigneeCommon Services Agency
InventorsPeter Simmonds, Shui-Wan Chan, Peng Lee Yap
Life of this patent
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Abstract
New styles of hepatitis C virus (HCV), referred to as HCV-3 and HCV-4, have been identified and sequenced. Antigenic regions of HCV-2, HCV-3 and HCV-4 polypeptides have been identified. Immunoassays for HCV and antibodies thereto are described, which allow more complete screening of blood samples for HCV, and allow HCV genotyping.
Claims18
An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of:
- a) a nucleotide sequence encoding an amino acid sequence set forth in SEQ ID NO:3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 15, 17, or 21;
- and, b) a nucleotide sequence set forth in SEQ ID NO:
- 12, 16, 18, 19, or 20.
An isolated polypeptide comprising an amino acid sequence set forth in SEQ ID NO:3, 4, 5, 6, 9, 13, 14, 15, 17, or 21.
An antibody that selectively binds to a polypeptide having the amino acid sequence set forth in SEQ ID NO:
- 1, 2, 7, 8, 10, or 11.
An immunoassay device that comprises a solid substrate having attached thereto a polypeptide comprising an amino acid sequence set forth in SEQ ID NO:9, 13, 14, 15, 17, or 21.
A fusion polypeptide comprising a first polypeptide operably linked to a heterologous protein or fragment, wherein said first polypeptide is selected from the group consisting of the polypeptides set forth in SEQ ID NOS:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 15, and 21.
A fusion polypeptide comprising at least two polypeptides selected from the group consisting of the polypeptides set forth in SEQ ID NOS:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 15, 17, and 21, wherein said at least two polypeptides are operably linked together.
A vaccine formulation comprising at least one polypeptide selected from the group consisting of the polypeptides set forth in SEQ ID NOS:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 15, 17, and 21.
A method for detecting HCV infection in a mammal, comprising:
- a) obtaining a blood sample from a mammal;
- b) isolating RNA from said blood sample;
- c) reverse transcribing said RNA into cDNA;
- d) performing PCR on said cDNA using sense and antisense primers specific for HCV, wherein said sense and antisense primers are selected from the group consisting of:
- 12 i) SEQ ID NO:24 and SEQ ID NO:23;
- ii) SEQ ID NO:25 and SEQ ID NO:22;
- iii) SEQ ID NO:27 and SEQ ID NO:26;
- iv) SEQ ID NO:51 and SEQ ID NO:50;
- v) SEQ ID NO:52 and SEQ ID NO:53;
- vi) SEQ ID NO:42 and SEQ ID NO:40;
- vii) SEQ ID NO:42 and SEQ ID NO:41;
- viii) SEQ ID NO:43 and SEQ ID NO:40;
- and, ix) SEQ ID NO:43 and SEQ ID NO:41;
- and,
- e) determining whether HCV infection was detected by analyzing the results from said PCR.
A method for typing HCV infection in a mammal, comprising:
- a) obtaining a blood sample from a mammal;
- b) isolating RNA from said blood sample;
- c) reverse transcribing said RNA into cDNA;
- d) performing PCR on said cDNA using an antisense primer common to all HCV types and a sense primer specific for HCV type 1, 2, 3, or 4, wherein said antisense primer is selected from the group consisting of SEQ ID NO:32 and SEQ ID NO:33, and wherein said sense primer is selected from the group consisting of;
- 13 i) SEQ ID NO:34;
- ii) SEQ ID NO:35;
- iii) SEQ ID NO:36;
- iv) SEQ ID NO:37;
- v) SEQ ID NO:38;
- and, vi) SEQ ID NO:39;
- and,
- e) determining what type of HCV infection was detected by analyzing the results from said PCR.
A method for typing HCV infection in a mammal, comprising:
- a) obtaining a blood sample from a mammal;
- b) isolating RNA from said blood sample;
- c) reverse transcribing said RNA into cDNA;
- d) performing PCR on said cDNA using a non-type-specific antisense primer and a sense primer specific for HCV types 1, 2, 3, or 4, wherein said antisense primer is selected from the group consisting of SEQ ID NO:22 and SEQ ID NO:23, and wherein said sense primer is selected from the group consisting of;
- 14 i) SEQ ID NO:44;
- ii) SEQ ID NO:45;
- iii) SEQ ID NO:46;
- iv) SEQ ID NO:47;
- v) SEQ ID NO:48;
- and, vi) SEQ ID NO:49;
- and,
- e) determining what type of HCV infection was detected by analyzing the results from said PCR.
A method for typing HCV infection in a mammal, comprising:
- a) obtaining a blood sample from a mammal;
- b) isolating RNA from said blood sample;
- c) reverse transcribing said RNA into cDNA;
- d) performing PCR on said cDNA using an antisense primer and a sense primer specific for HCV type 3, wherein said sense and antisense primers are selected from the group consisting of:
- 15 i) SEQ ID NO:30 and SEQ ID NO:28;
- and, ii) SEQ ID NO:31 and SEQ ID NO:29;
- and,
- e) determining what type of HCV infection was detected by analyzing the results from said PCR.
A method for typing HCV infection in a mammal, comprising:
- a) obtaining a blood sample from a mammal;
- b) isolating nucleic acid from said sample;
- c) digesting said nucleic acid separately with ScrF1, HaeIII/RsaI, and Hinf1 to produce three samples of digested nucleic acid;
- d) analyzing the restriction pattern of said samples to differentiate between type 1, type 2, type 3, and type 4 HCV infection.
A method for detecting HCV infection in a mammal, comprising:
- a) obtaining a blood sample from said mammal;
- b) contacting said blood sample with at least one polypeptide selected from the group consisting of the polypeptides set forth in SEQ ID NOS:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 14, 15, 17, and 21;
- and,
- c) detecting if an antibody present in said blood sample is bound to said polypeptide.
Dependencies are inferred from the claim text.