Clinical evaluation of oligonucleotide microarrays for the detection of HBV mutants associated with lamivudine resistance.
OBJECTIVE
Long-term lamivudine administration in hepatitis B virus (HBV)-infected patients induces the emergence of HBV mutants with lamivudine resistance. The aim of the present study was to evaluate the clinical application of an oligonucleotide microarray in detecting HBV mutants associated with lamivudine resistance.


METHODS
947 HBV DNA-positive sera from: 388 patients receiving lamivudine treatment, 559 chronic hepatitis B patients not receiving lamivudine treatment, and 359 from HBV DNA-negative controls, were assayed for HBV mutations using the oligonucleotide microarray. Furthermore, follow-up studies were performed using 255 clinical samples from 51 patients treated with lamivudine at various periods. The results were compared with sequencing and real-time polymerase chain reaction (PCR).


RESULTS
The HBV DNA polymerase Tyr-Met-Asp-Asp motif (YMDD) mutation was detected in all 388 samples containing lamivudine-resistant mutations identified by microarray. For the codons rt180, rt204 and rt207, the agreements between the microarray and sequencing data are 96.6, 98.5 and 100%, respectively. Two previously unreported mutants were also found in those samples. In the 947 samples collected from different patients, which were detected positive for HBV DNA by quantitative PCR, all but three weak-positive samples were positive by the microarray, demonstrating an agreement of 99.7%. In all the positive samples, mutations could be detected in the relevant loci of HBV DNA polymerase with lamivudine resistance. All of the 359 HBsAg-negative samples were shown to be negative for HBV DNA using the microarray method. Follow-up detection of the clinical samples from 51 patients treated with lamivudine demonstrated that the microarray method was able to detect mutations in mixed viruses that were infecting prior to sequencing.


CONCLUSIONS
The oligonucleotide microarray can be conveniently utilized to detect mutant HBV in clinical serum samples.
Chen
Li-Yan
LY
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Huang
Jian
J
Zhang
Xin-Ping
XP
Qiao
Ping
P
Zhang
Wen
W
Yang
Ning-Min
NM
Liu
Hong-Jing
HJ
Geng
Yong-Yao
YY
Qiu
Jian-Ming
JM
Wang
Sheng-Qi
SQ
D000998:Antiviral Agents / Q000008:administration & dosage* / Q000009:adverse effects; D016022:Case-Control Studies; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D024882:Drug Resistance, Viral / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000097:blood / Q000188:drug therapy* / Q000821:virology; D006801:Humans; D019259:Lamivudine / Q000008:administration & dosage* / Q000009:adverse effects; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D012680:Sensitivity and Specificity
D000998:Antiviral Agents; D004279:DNA, Viral; D019259:Lamivudine