Viral variants and methods for detecting same
AssigneeWestern Health Care Network
InventorsStephen A Locarnini, Angeline I. Bartholomeusz, Thein T. Aye, Robert A. de Man
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Abstract
The present invention relates generally to viral variants exhibiting reduced sensitivity to particular agents and/or reduced interactivity with immunological reagents. More particularly, the present invention is directed to hepatitis B variants exhibiting complete or partial resistance to nucleoside analogues and/or reduced interactivity with antibodies to viral surface components. The present invention further contemplates assays for detecting such viral variants which assays are useful in monitoring anti-viral therapeutic regimes.
Claims25
An isolated HBV mutant, comprising a mutation in the gene encoding the HBV DNA polymerase resulting in decreased sensitivity to a nucleoside analogue compared to a wild-type HBV, wherein said mutation results in at least one amino acid addition, substitution, and/or deletion in the B domain corresponding to amino acid residues 495-535 of a wild-type HBV polymerase, and said mutant contains an unmutated YMDD motif in the C domain.
An isolated HBV mutant, comprising a mutation in the gene encoding the HBV DNA polymerase resulting in decreased sensitivity to a nucleoside analogue compared to a wild-type HBV, wherein said mutation results in at least one amino acid addition, substitution, and/or deletion in the B domain corresponding to amino acid residues 505-535 of a wild-type HBV polymerase, and said mutant contains an unmutated YMDD motif in the C domain.
An isolated HBV mutant, comprising a mutation in the gene encoding the HBV DNA polymerase resulting in decreased sensitivity to a nucleoside analogue compared to a wild-type HBV, wherein said mutation results in at least one amino acid addition, substitution, and/or deletion in the B domain corresponding to amino acid residues 505-529 of a wild-type HBV polymerase, and said mutant contains an unmutated YMDD motif in the C domain.
An isolated HBV mutant exhibiting, relative to an isolated wild-type HBV, reduced sensitivity to a nucleoside analogue and reduced interactivity to an antibody to a wild-type HBV surface antigen, said HBV mutant comprising at least one of:
- (i) a nucleotide sequence of its genome as set forth in SEQ ID NO:17 or a sequence having at least 60% similarity thereto;
- (ii) a nucleotide sequence capable of hybridising to SEQ ID NO:17 under low stringency conditions at 42° C.;
- (iii) a mutation in an overlapping portion of open reading frames for DNA polymerase and HBV surface antigen;
- and
- (iv) a mutation in a region corresponding to amino acids 118 to 169 and/or 169 to 207 of HBV surface antigen,
- wherein said mutant contains a Trp/Arg499Glu amino acid substitution in the DNA polymerase and an Asp144Glu and Gly145Arg amino acid substitution in the surface antigen.
An isolated HBV mutant exhibiting, relative to an isolated wild-type HBV, reduced sensitivity to a nucleoside analogue, said mutant comprising at least one mutation in its genome wherein said at least one mutation produces at least one amino acid substitution in the DNA polymerase selected from the group consisting of Trp/Arg499Glu, Phe512Leu and Val519Leu, said amino acid substitution in the DNA polymerase resulting in a concurrent amino acid substitution in the overlapping open reading frame of the HBV surface antigen, and said mutant contains an unmutated YMDD motif in the C domain.
An isolated HBV mutant exhibiting, relative to an isolated wild-type HBV, reduced sensitivity to a nucleoside analogue, said mutant comprising at least one mutation in its genome wherein said at least one mutation produces at least one amino acid substitution in the DNA polymerase selected from the group consisting of Trp/Arg499Glu, Phe512Leu, Val519Leu and Ser559Thr, said amino acid substitution in the DNA polymerase resulting in a concurrent amino acid substitution in the overlapping open reading frame of the HBV surface antigen, and said mutant contains an unmutated YMDD motif in the C domain.
A method for determining the potential for an HBV to exhibit, relative to an isolated wild-type HBV, reduced sensitivity to at least one of lamivudine, penciclovir and famciclovir, said method comprising isolating DNA or corresponding mRNA from said HBV and screening for a mutation in a nucleotide sequence encoding the B domain of HBV polymerase. corresponding to amino acid residues 495-535 of a wild-type HBV polymerase, with said mutation resulting in at least one amino acid substitution, deletion and/or addition in said B domain,
- wherein the presence of such a mutation is an indication of the potential of reduced sensitivity of said HBV to at least one of lamivudine, penciclovir and famciclovir.
A method for determining the potential for an HBV to exhibit, relative to an isolated wild-type HBV, reduced sensitivity to at least one of lamivudine, penciclovir and famciclovir, said method comprising isolating DNA or corresponding mRNA from said HBV and screening for a mutation in a nucleotide sequence encoding the B domain of HBV polymerase corresponding to amino acid residues 505-535 of a wild-type HBV polymerase, with said mutation resulting in at least one amino acid substitution, deletion and/or addition in said B domain,
- wherein the presence of such a mutation is an indication of the potential of reduced sensitivity of said HBV to at least one of lamivudine, penciclovir and famciclovir.
A method for determining the potential for an HBV to exhibit, relative to an isolated wild-type HBV, reduced sensitivity to at least one of lamivudine, penciclovir and famciclovir, said method comprising isolating DNA or corresponding mRNA from said HBV and screening for a mutation in a nucleotide sequence encoding the B domain of HBV polymerase corresponding to amino acid residues 505-529 of a wild-type HBV polymerase, with said mutation resulting in at least one amino acid substitution, deletion and/or addition in said B domain,
- wherein the presence of such a mutation is an indication of the potential of reduced sensitivity of said HBV to at least one of lamivudine, penciclovir and famciclovir.
A method for determining the potential for an HBV to exhibit, relative to an isolated wild-type HBV, reduced sensitivity to at least one of lamivudine, penciclovir and famciclovir, said method comprising isolating DNA or corresponding mRNA from said HBV and screening for a mutation in a nucleotide sequence encoding HBV DNA polymerase, wherein the screening detects at least one mutation selected from the group consisting of Arg/Trp499Glu, Phe512Leu, Val 519Leu, Pro523Leu, Leu526Met, Thr530Ser, Ile533Leu, Met550Val and Met550Ile,
- such that when said at least one amino substitution in the DNA polymerase is Met550Val or Met550Ile, said method detects at least one amino acid substitution other than Phe512Leu, Leu526Met or Val553Ile,
- wherein the presence of such a mutation is an indication of the potential of reduced sensitivity of said HBV to at least one of lamivudine, penciclovir and famciclovir.
A method for determining the potential for an HBV to exhibit, relative to an isolated wild-type HBV, reduced sensitivity to at least one of penciclovir and famciclovir, said method comprising isolating DNA or corresponding mRNA from said HBV and screening for a mutation in a nucleotide sequence encoding HBV DNA polymerase, wherein the screening detects at least one mutation selected from the group consisting of Arg/Trp499Glu, Phe512Leu, Val 519Leu, Pro523Leu, Leu526Met, Thr530Ser, Ile533Leu, Met550Val and Met550Ile,
- wherein the presence of such a mutation is an indication of the potential of reduced sensitivity of said HBV to at least one of penciclovir and famciclovir.
An isolated Hepadnavirus mutant, comprising a mutation in the gene encoding the DNA polymerase, resulting in decreased sensitivity to a nucleoside analogue compared to a wild-type Hepadnavirus, wherein said mutation results in at least one amino acid addition, substitution, and/or deletion in the B domain corresponding to amino acid residues 495-535 of a wild-type HBV polymerase, and said mutant contains an unmutated YMDD motif in the C domain.
An isolated Hepadnavirus mutant, comprising a mutation in the gene encoding the DNA polymerase, resulting in decreased sensitivity to a nucleoside analogue compared to a wild-type Hepadnavirus, wherein said mutation results in at least one amino acid addition, substitution, and/or deletion in the B domain corresponding to amino acid residues 505-535 of a wild-type HBV polymerase, and said mutant contains an unmutated YMDD motif in the C domain.
An isolated Hepadnavirus mutant, comprising a mutation in the gene encoding the DNA polymerase, resulting in decreased sensitivity to a nucleoside analogue compared to a wild-type Hepadnavirus, wherein said mutation results in at least one amino acid addition, substitution, and/or deletion in the B domain corresponding to amino acid residues 505-529 of a wild-type HBV polymerase, and said mutant contains an unmutated YMDD motif in the C domain.
A method for determining the potential for an HBV to exhibit, relative to an isolated wild-type HBV, reduced sensitivity to an anti-viral agent, said method comprising isolating DNA or corresponding mRNA from said HBV and screening for a mutation in a nucleotide sequence encoding the B domain of HBV polymerase corresponding to amino acid residues 495-535 of a wild-type HBV polymerase, with said mutation resulting in at least one amino acid substitution, deletion and/or addition in said B domain,
- wherein the presence of such a mutation is an indication of the potential of reduced sensitivity of said HBV to an anti-viral agent.
Dependencies are inferred from the claim text.