Mutation detection in the drug-resistant hepatitis B virus polymerase gene using nanostructured reverse micelles.
The emergence of drug-resistant hepatitis B virus (HBV) has been reported in patients with prolonged administration of lamivudine, which is a potent drug for the prevention of HBV infection. Lamivudine-resistant HBV has several types of mutations at the YMDD motif of its DNA polymerase. We successfully demonstrated that monitoring the hybridization behavior in nanostructured reverse micelles enables us to detect single nucleotide polymorphisms (SNPs). With the aid of reverse micelles, a model 40-mer oligonucleotide containing a single-base substitution was clearly distinguished from the normal, complementary oligonucleotide. In addition, we extended this technique to a high-throughput analysis. The results obtained with a 96-well micro-plate reader indicated the possibility of SNPs detection toward multiple samples of patients.
Park
Lian-Chun
LC
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
Maruyama
Tatsuo
T
Kamiya
Noriho
N
Goto
Masahiro
M
Kuma
Hiroyuki
H
Hamasaki
Naotaka
N
D019380:Anti-HIV Agents / Q000494:pharmacology; D006515:Hepatitis B virus / Q000201:enzymology* / Q000235:genetics*; D017403:In Situ Hybridization; D007202:Indicators and Reagents; D019259:Lamivudine / Q000494:pharmacology; D008823:Micelles; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000235:genetics*; D013056:Spectrophotometry, Ultraviolet
D019380:Anti-HIV Agents; D007202:Indicators and Reagents; D008823:Micelles; D009841:Oligonucleotides; D019259:Lamivudine; D012194:RNA-Directed DNA Polymerase