title: Integrated chip electrophoresis and magnetic particle isolation used for detection of hepatitis B virus oligonucleotides.
abstract: Rapid and sensitive detection is a key step in the effective and early response to the global hazard of various viral diseases. In this study, an integrated isolation of hepatitis B virus (HBV)-specific DNA fragment by magnetic nanoparticles (MNPs) and its immediate analysis by microchip CGE was performed. Microfluidic CE chip was used to accommodate the complete process of viral DNA isolation by MNPs including hybridization and thermal denaturation followed by CE separation. Beforehand, calibration curves of HBV fragments were constructed. For isolation by MNPs, specific streptavidin-biotin interaction was used to bind complementary HBV fragment to magnetic particles. After analysis of isolated HBV by regular MNPs method, innovative approach was performed. The commercial CE chip (Bio-rad) was successfully used to execute HBV fragment isolation. Detection using LIF with detection limit of 1 ng/mL was accomplished.
authors.affiliation: Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Brno, Czech Republic.
authors.forename: Marketa
authors.initials: M
authors.lastname: Vaculovicova
authors.forename: Kristyna
authors.initials: K
authors.lastname: Smerkova
authors.forename: Jiri
authors.initials: J
authors.lastname: Sedlacek
authors.forename: Jiri
authors.initials: J
authors.lastname: Vyslouzil
authors.forename: Jaromir
authors.initials: J
authors.lastname: Hubalek
authors.forename: Rene
authors.initials: R
authors.lastname: Kizek
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
mesh_terms: D004279:DNA, Viral / Q000302:isolation & purification*; D046209:Electrophoresis, Microchip / Q000295:instrumentation*; D006509:Hepatitis B / Q000175:diagnosis; D006515:Hepatitis B virus / Q000302:isolation & purification*; D006801:Humans; D057230:Limit of Detection; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D009841:Oligonucleotides / Q000302:isolation & purification*
chemical_list: D004279:DNA, Viral; D058185:Magnetite Nanoparticles; D009841:Oligonucleotides