Analytical Sciences

Mutation Detection in the Drug-Resistant Hepatitis B Virus Polymerase Gene Using Nanostructured Reverse Micelles

Lian-Chun PARK,* Tatsuo MARUYAMA,* Noriho KAMIYA,* Masahiro GOTO,*^,** Hiroyuki KUMA,*** and Naotaka HAMASAKI***

*Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan
**PRESTO, JST (Japan Science and Technology Corporation)
***Department of Clinical Chemistry and Laboratory Medicine, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan

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.

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