title: Chemiluminescence-imaging detection of DNA on a solid-phase membrane by using a peroxidase-labeled macromolecular probe.
abstract: We have developed a novel method for sensitive chemiluminescence (CL)-imaging detection of DNA by using a macromolecular probe synthesized by attaching multiple molecules of horseradish peroxidase (HRP) and biotin in dextran backbone. The probe formed a macromolecular assembly by binding to streptavidin which specifically recognized biotinylated complementary DNA, which was hybridized to a target DNA on a solid-phase membrane. This methodology was applied to CL-imaging detection of a synthetic telomere DNA (TTAGGG)10 and human telomere DNA by using the CL probe comprising of dextranT2000 (MW=ca. 2000kDa) bonded to approximately 42 molecules of HRP and 210 molecules of biotin. The human telomere DNA in a small number of buccal mucous cells (ca. 70 cell numbers) of cheek tissue was quantitatively determined by the proposed CL detection method that afforded approximately 10 times higher sensitivity than that of the conventional CL method using commercially available HRP-avidin probe.
authors.affiliation: Department of Pharmaceutical Sciences, North South University, Basundhara, Dhaka 1229, Bangladesh; Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-Machi 1-14, Nagasaki 852-8521, Japan.
authors.forename: Md Golam
authors.initials: MG
authors.lastname: Azam
authors.affiliation: Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-Machi 1-14, Nagasaki 852-8521, Japan.
authors.forename: Mutsumi
authors.initials: M
authors.lastname: Yamasuji
authors.affiliation: Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-Machi 1-14, Nagasaki 852-8521, Japan; Department of Chemical Organic Technology and Petrochemistry, Faculty of Chemistry, Silesian University of Technology, Krzywoustego 4, 44-100 Gliwice, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Krawczyk
authors.affiliation: Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-Machi 1-14, Nagasaki 852-8521, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Shibata
authors.affiliation: Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-Machi 1-14, Nagasaki 852-8521, Japan.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Kabashima
authors.affiliation: Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Bunkyo-Machi 1-14, Nagasaki 852-8521, Japan. Electronic address: ms-kai@nagasaki-u.ac.jp.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Kai
mesh_terms: D000328:Adult; D001360:Avidin / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D015139:Blotting, Southern; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis*; D003911:Dextrans / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D046911:Macromolecular Substances / Q000737:chemistry*; D009061:Mouth Mucosa / Q000737:chemistry*; D009693:Nucleic Acid Hybridization / Q000379:methods; D016615:Telomere / Q000737:chemistry*
chemical_list: D003911:Dextrans; D046911:Macromolecular Substances; D001360:Avidin; D001710:Biotin; D004247:DNA; D006735:Horseradish Peroxidase
keywords: Chemiluminescence detection; Horseradish peroxidase; Macromolecular probe; Solid-phase membrane; Telomere DNA