Establishment of a new quantitative detection approach to adefovir-resistant HBV and its clinical application.
OBJECTIVE
To establish the more feasible and sensitive assessment approach to the detection of adefovir (ADV) resistance-associated hepatitis B virus (HBV) quasispecies.


METHODS
Based on the characteristics of rtA181V/T and rtN236T mutations, a new approach based on real-time fluorescent quantitative polymerase chain reaction (RT-PCR) was established for the detection of ADV-resistant HBV quasispecies, total HBV DNA, rtA181 and rtN236 mutations in blood samples from 32 chronic hepatitis B (CHB) patients with unsatisfactory curative effect on ADV and compared with routine HBV DNA sequencing.


RESULTS
Both the sensitivity and specificity of this new detection approach to ADV-resistant HBV quasispecies were 100%, which were much higher than those of direct HBV DNA sequencing. The approach was able to detect 0.1% of mutated strains in a total plasmid population. Among the 32 clinical patients, single rtA181 and rtN236T mutation and double rtA181T and rtN236T mutations were detected in 20 and 8, respectively, while ADV-resistant mutations in 6 (including, rtA181V/T mutation alone in 5 patients) and no associated mutations in 26.


CONCLUSIONS
This new approach is more feasible and efficient to detect ADV-resistant mutants of HBV and ADV-resistant mutations before and during ADV treatment with a specificity of 100% and a sensitivity of 100%.
Zhao
Wei-Feng
WF
Shanghai Laboratory of Disease and Health Genomics, Chinese National Human Genome Center, Shanghai 201203, China.
Shao
You-Lin
YL
Chen
Liang-Yun
LY
Wu
Jin-Hua
JH
Zhu
Yi-Ling
YL
Gan
Jian-He
JH
Xiong
Hui
H
D000225:Adenine / Q000031:analogs & derivatives*; D000328:Adult; D000998:Antiviral Agents; D004279:DNA, Viral / Q000235:genetics; D024882:Drug Resistance, Viral; D005260:Female; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000188:drug therapy / Q000821:virology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D063065:Organophosphonates; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D055815:Young Adult
D000998:Antiviral Agents; D004279:DNA, Viral; D063065:Organophosphonates; C053001:adefovir; D000225:Adenine