title: Design of a sandwich-mode amperometric biosensor for detection of PML/RARα fusion gene using locked nucleic acids on gold electrode.
abstract: In this study, a novel DNA electrochemical probe (locked nucleic acid, LNA) was designed and involved in constructing an electrochemical DNA biosensor for detection of promyelocytic leukemia/retinoic acid receptor alpha (PML/RARα) fusion gene in acute promyelocytic leukemia for the first time. This biosensor was based on a 'sandwich' sensing mode, which involved a pair of LNA probes (capture probe immobilized at electrode surface and biotinyl reporter probe as an affinity tag for streptavidin-horseradish peroxidase (streptavidin-HRP). Since biotin can be connected with streptavidin-HRP, this biosensor offered an enzymatically amplified electrochemical current signal for the detection of target DNA. In the simple hybridization system, DNA fragment with its complementary DNA fragment was evidenced by amperometric detection, with a detection limit of 74 fM and a linear response range of 0.1-10 pM for synthetic PML/RARα fusion gene in acute promyelocytic leukemia (APL). Otherwise, the biosensor showed an excellent specificity to distinguish the complementary sequence and different mismatch sequences. The new pattern also exhibited high sensitivity and selectivity in mixed hybridization system.
authors.affiliation: Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
authors.forename: Zhouliang
authors.initials: Z
authors.lastname: Sun
authors.forename: Meijuan
authors.initials: M
authors.lastname: Feng
authors.forename: Ailin
authors.initials: A
authors.lastname: Liu
authors.forename: Shuyu
authors.initials: S
authors.lastname: Yang
authors.forename: Yuanzhong
authors.initials: Y
authors.lastname: Chen
authors.forename: Xinhua
authors.initials: X
authors.lastname: Lin
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods / Q000706:statistics & numerical data; D015342:DNA Probes / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D055664:Electrochemical Techniques; D004566:Electrodes; D004867:Equipment Design; D006046:Gold; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000175:diagnosis / Q000235:genetics; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics*; D015514:Oncogene Proteins, Fusion / Q000235:genetics*
chemical_list: D015342:DNA Probes; D004273:DNA, Neoplasm; D009841:Oligonucleotides; D015514:Oncogene Proteins, Fusion; C477371:locked nucleic acid; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein; D006046:Gold