HBV drug resistance methods
New polymorphisms in the nucleic acid sequences of the DNA polymerase/reverse transcriptase open reading frame and viral surface antigen open reading frame of the hepatitis B virus are reported. In particular, the present invention relates to the mutation YMDD→YSDD in the HBV reverse transcriptase domain and to the W196V mutation in the small HBV viral surface antigen. Said polymorphisms are affecting the detection of drug resistance mutations by genotypic methods and diagnostic kits based thereon. The present invention relates to methods and diagnostic kits for detection of a HBV virus comprising said nucleic acid polymorphisms. In particular, those methods utilizing oligonucleotides capable of hybridizing to said HBV nucleic acid polymorphisms are envisaged.
1. A diagnostic kit for detecting resistance to lamivudine or a combination of antiviral drugs comprising lamivudine of an HBV virus present in a biological sample comprising an oligonucleotide capable of discriminating, in said HBV polynucleic acid, a codon at the position corresponding to position 204 of sequence P7RT (position 108 of SEQ ID NO:4) encoding a serine from a codon at the position corresponding to position 204 of sequence P7RT (position 108 of SEQ ID NO:4) encoding a methionine, valine or isoleucine.
2. The diagnostic kit according to claim 1 further comprising an oligonucleotide capable of discriminating, in said HBV polynucleic acid, a codon at the position corresponding to position 180 of sequence P7RT (position 84 of SEQ ID NO:4) encoding a methionine from a codon at the position corresponding to position 180 of sequence P7RT (position 84 of SEQ ID NO:4) encoding a leucine.
3. The diagnostic kit according to claim 1 wherein said oligonucleotide is attached or immobilized to a solid support.
4. The diagnostic kit according to claim 2 wherein at least one of
said oligonucleotide capable of discriminating, in said HBV polynucleic acid, a codon at the position corresponding to position 204 of sequence P7RT (position 108 of SEQ ID NO:4) encoding a serine from a codon at the position corresponding to position 204 of sequence P7RT (position 108 of SEQ ID NO:4) encoding a methionine, valine or isoleucine is attached or immobilized to a solid support and
said oligonucleotide capable of discriminating, in said HBV polynucleic acid, a codon 180 encoding a methionine from a codon 180 encoding a leucine is attached or immobilized to a solid support.