分析测试学报2011，Vol.30Issue(5)：492-497,6.DOI:10.3969/j.issn.1004-4957.2011.05.004
检测急性早幼粒细胞白血病PML/RARα融合基因的电化学传感器研制
Development of Electrochemical Biosensor for Detection of PML/RARα Fusion Gene in Acute Promyelocytic Leukemia
摘要
Abstract
A novel DNA electrochemical probe (locked nucleic acid, LNA) was designed and involved in constructing an electrochemical DNA biosensor for the detection of PML/RARα fusion gene in acute promyelocytic leukemia(APL). This biosensor was based on a “sandwich” detection strategy, which involved a pair of LNA probes, e.g. hairpin capture probe and reporter probe. Streptavi-din -HRP was bound to biotin labeled at the end of reporter probe via streptavidin -biotin affinity binding. In the presence of hydrogen peroxide( H2O2 ), HRP catalyzed the oxidation of the substrate 3, 3′, 5, 5′-tetramethylbenzidene (TMB) to offer an enzymatically amplified electrochemical current signal for the detection of target DNA. This sensor was applied in the direct quantitative detection of synthetic PML/RARα fusion gene in PBS buffer. The results indicated that the biosensor showed an excellent specificity to distinguish the complementary sequence and different mismatch sequences. A linear relationship between the amperometric signal and the target concentration was obtained in the range of 1.0 × 10-11 - 1.6 × 10-10 mol/L with a detection limit of 1.0 × 10-13 mol/L. In addition, the biosensor was used for the determination of PML/RARα fusion gene in human serum samples without dilution with high sensitivity, selectivity and good repeatability. This method would be expected to use in real sample for further solving the actural problems of early diagnosis and prognosis monitoring of APL.关键词
电化学传感器/"三明治"结构/发夹结构DNA探针/锁核酸/PML/RARα融合基因Key words
electrochemical biosensors/ sandwich structure/ hairpin DNA probe/ locked nucleic acid/ PML/RARα fusion gene分类
化学化工引用本文复制引用
冯美娟,雷云,王昆,陈元仲,李光文,罗红斌,林新华..检测急性早幼粒细胞白血病PML/RARα融合基因的电化学传感器研制[J].分析测试学报,2011,30(5):492-497,6.基金项目
863计划资助项目(2008AA02Z433) （2008AA02Z433）
福建省高校产学研科技重点项目(2010Y4003) （2010Y4003）
国家自然科学基金资助项目(20805006,20975021) （20805006,20975021）
福建省自然科学基金资助项目(2010J05019) （2010J05019）
