Variants of hepatitis b virus with resistance to anti-viral nucleoside agents and applications thereof

A method for determining the potential for an HBV to exhibit reduced sensitivity to one or more nucleoside or nucleotide analogs selected from the group consisting of ADV;

• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• or ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV said method comprising isolating DNA or corresponding mRNA from the HBV and screening for a mutation in HBV nucleotide sequence and associated with resistance or decreased sensitivity to ADV;
• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV with the proviso that the HBV variant is not resistant to ETV alone or ETV and LMV alone.

A composition comprising a variant HBV resistant to one or more nucleoside or nucleotide analogs selected from the group consisting of ADV;

• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• or ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV, and one or more pharmaceutically acceptable carriers and/or diluents with the proviso that the HBV variant does not exhibit resistance to ETV alone or ETV or LMV alone.

A method for determining whether an HBV strain exhibits reduced sensitivity to a nucleoside or nucleotide analog said method comprising isolating DNA or corresponding mRNA from the HBV and screening for a mutation in the nucleotide sequence encoding the DNA polymerase wherein the presence of a mutation in the rt region selected from a codon selected from the group consisting of rtN53, rtN53, rtY54, rtL180, rtT181, rtT184, rtM204, rtN236, rtY124, rtH126, rtT128, rtS135, rtL180, rtS202, rtM204, rtH248, rtI53, rtS54, rtN122, rtM145, rtL180, rtM204, rtM250, rtN53, rtS85, rtS116, rtD134, rtN139, rtQ149, rtA181, rtS219, rtI233, rtN236 and rtM250 is indicative of a variant which exhibits a decreased sensitivity to one or more nucleoside or nucleotide analogs selected from the group consisting of ADV;

• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV with the proviso that the HBV variant does not exhibit resistance to ETV alone or ETV or LMV alone.

A method for determining whether an HBV strain exhibits reduced sensitivity to a nucleoside or nucleotide analog said method comprising screening for a mutation in the nucleotide sequence encoding the envelope genes (s) wherein the presence of a mutation in the S gene at a codon selected from the group consisting of sT45, sL173, sL175, sI195, sT118, sP120, sP127, sI195, sT114, sI195, sS204, sI208, sS210, sV14, sG130, sM133, sW172, sS204 and sS210 is indicative of a variant which exhibits a decreased sensitivity to one or more nucleoside or nucleotide analogs selected from the group consisting of ADV;

• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV with the proviso that the HBV variant is not resistant to ETV alone or ETV and LMV alone.

A method for detecting a variant HBV exhibiting an altered immunological profile, said method comprising isolating an HBV from a subject exposed to a nucleoside or nucleotide analog or combination of analogs selected from the listed consisting of ADV;

• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV said method comprising contacting said HBV with a panel of one or more antibodies to a surface antigen and screening for any change in binding affinity or binding spectrum with the proviso that the HBV variant does not exhibit resistance to ETV alone or ETV or LMV alone.

A method for detecting a variant HBV exhibiting an altered immunological profile said method comprising isolating a serum sample from a subject exposed to one or more nucleoside or nucleotide analog selected from the group consisting of ADV;

• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV, and then contacting the serum with a panel of HBV surface antigens or antibody-binding fragments thereof and screening for any change in binding affinity or binding spectrum with the proviso that the HBV variant does not exhibit resistance to ETV alone or ETV or LMV alone.

A vaccine against HBV infection wherein the HBV is resistant to one or more nucleoside or nucleotide analogs selected from the group consisting of ADV;

• LMV;
• TFV;
• FTC;
• ADV and LMV;
• ADV and TFV;
• LMV and TFV;
• FTC and ADV;
• FTC and TFV;
• FTC and LMV;
• ETV and ADV;
• ETV and FTC;
• ETV and TFV;
• ADV and LMV and TFV;
• ADV and FTC and TFV;
• TFV and FTC and LMV;
• ADV and LMV and ETV;
• ADV and ETV and TFV;
• ETV and LMV and TFV;
• ADV and LMV and FTC;
• ADV and FTC and LMV and TFV;
• ETV and FTC and LMV and TFV;
• ADV and ETV and LMV and TFV;
• ADV and FTC and ETV and TFV;
• ADV and FTC and LMV and ETV;
• and ADV and FTC and LMV and TFV and ETV said vaccine comprising a HBV vaccine comprising a mutation at a codon selected from the group consisting of rtN53, rtY54, rtL180, rtT181, rtT184, rtM204, rtN236, rtY124, rtH126, rtT128, rtS135, rtL180, rtS202, rtM204, rtH248, rtI53, rtS54, rtN122, rtM145, rtL180, rtM204, rtM250, rtN53, rtS85, rtS116, rtD134, rtN139, rtQ149, rtA181, rtS219, rtI233, rtN236 and rtM250 with the proviso that the HBV variant does not exhibit resistance to ETV alone or ETV or LMV alone.