Drug resistance motifs HBV Phenotypic resistance Clinical resistance

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耐药分析 • 耐药基序 (Drug resistance motifs) • HBV表型耐药 (Phenotypic resistance) • 临床耐药 (Clinical resistance)

耐药分析 • 耐药基序 (Drug resistance motifs) • HBV表型耐药 (Phenotypic resistance) • 临床耐药 (Clinical resistance) 三者相关,但不完全等同

Lamivudine and wild-type HBV(一) Y Nucleotides high affinity ss (-) DNA M D D

Lamivudine and wild-type HBV(一) Y Nucleotides high affinity ss (-) DNA M D D HBV polymerase (wild-type)

Lamivudine and wild-type HBV(二) Lamivudine Y Nucleotides high affinity inhibition ss (-) DNA M

Lamivudine and wild-type HBV(二) Lamivudine Y Nucleotides high affinity inhibition ss (-) DNA M D D HBV polymerase (wild-type)

Lamivudine and YMDD variant HBV Lamivudine Y weak inhibition Nucleotides reduced affinity V/I ss

Lamivudine and YMDD variant HBV Lamivudine Y weak inhibition Nucleotides reduced affinity V/I ss (-) DNA D D HBV polymerase (variant)

耐药变异的检测方法 • Sequences analysis • Line probe assay • PCR-RFLP analysis (Restriction fragment length

耐药变异的检测方法 • Sequences analysis • Line probe assay • PCR-RFLP analysis (Restriction fragment length polymorphism) • Real-time PCR • Molecular beacons • DNA chip

Light. Cycler Detection Formats l Mutation Analysis Mismatch Perfect Match Temperature low medium high

Light. Cycler Detection Formats l Mutation Analysis Mismatch Perfect Match Temperature low medium high

YMDD基序变异发生后的临床影响 Incidence of YMDD variant HBV l Incidence of detectable serum YMDD variant HBV

YMDD基序变异发生后的临床影响 Incidence of YMDD variant HBV l Incidence of detectable serum YMDD variant HBV in HBe. Ag+ve patients after lamivudine therapy for: – – l 1 yr = 24% (Lai et al, 2001) 2 yr = 38% (Liaw et al, 2000) 3 yr = 49% (Leung et al, 1999) 4 yr = 67% (Chang et al, 2000) Emergence of YMDD variants does not necessarily equate to clinical resistance

YMDD变异检测结果 编号 治疗前 YMDD YIDD YVDD 治疗 48周 变异株百 分比(%) YMDD YIDD YVDD 变异株百

YMDD变异检测结果 编号 治疗前 YMDD YIDD YVDD 治疗 48周 变异株百 分比(%) YMDD YIDD YVDD 变异株百 分比(%) 1 17 1 1 10. 5 0 1 23 100 2 19 3 0 13. 6 0 1 18 100 3 21 0 2 8. 7 6 0 11 65 4 21 0 0 20 0 100 5 17 0 3 15 20 0 张欣欣等 瑞金医院

Country Method Number of Patients Percents of Detection of YMDD mutant in untreated patients

Country Method Number of Patients Percents of Detection of YMDD mutant in untreated patients China Clone Analysis 5 80% (4/5) Japan 1 PCR –ELISA & mini-sequence method 18 28% (5/18) Japan 2 Peptide nucleic acid (PNA) mediated polymerase chain reaction clamping 18 22% (4/18) Germany Line- probeassay 86 • Of the patients who developed resistance, 32% had Japan 1: Satsuki Kobayashi, Tatsuya Ide, Michio Sata YMDD mutants pretreatment. • Of those who did not develop resistance, 59% had YMDD mutants pretreatment. • Therefore, YMDD pre-treated doesn’t predict treatment failure. Japan 2: Toshihiko Kirishima, Takeshi Okanoue, Yukiko Daimon, et al.

Inhibition of lamivudine-Resistant HBV by Adefovir in Cell Culture Assay Mutation Wide type V

Inhibition of lamivudine-Resistant HBV by Adefovir in Cell Culture Assay Mutation Wide type V 521 L L 528 M M 552 I L 528 M + M 552 V V 521 L + L 528 M + M 552 V Fold Increase in IC 50 Lamivudine Adefovir 1 1 1. 4 1. 9 3. 5 1. 9 380 3. 2 2080 2. 5 1666 0. 7 L 528 M & V 521 L: compensatory mutations enhancing replication fitness

Viral Load Suppression in Patients Treated with ADV for Up to 72 Weeks Median

Viral Load Suppression in Patients Treated with ADV for Up to 72 Weeks Median Decrease in Duration of ADV Therapy ) No. of patients* HBV DNA (Log 10 c/ml 24 weeks 106 - 3. 6 48 weeks 46 - 4. 4 72 weeks 14 - 4. 7 Genotyped patients with both baseline and post-treatment HBV DNA

患者用拉米夫定前后的病情 演变 日达仙 1. 6 mg TIW ALT 拉米夫定 100 mg HBe. Ag+ HBe.

患者用拉米夫定前后的病情 演变 日达仙 1. 6 mg TIW ALT 拉米夫定 100 mg HBe. Ag+ HBe. Ab- DNA 干扰素 HBe. Ag- HBe. Ab+ 108 7 HBs. Ag- 10 106 105 104 103 102 男性,36岁