An Electrochemical DNA Biosensor for the Detection of the Apa I Polymorphism in the Vitamin D Receptor Gene Using Meldola's Blue as a Hybridization Indicator
An Electrochemical DNA Biosensor for the Detection of the Apa I Polymorphism in the Vitamin D Receptor Gene Using Meldola's Blue as a Hybridization Indicator
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DOI：
10.1002/elan.200900405
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发表时间：
2010-03-01
期刊：
影响因子：
3
通讯作者：
Ozsoz, Mehmet
中科院分区：
文献类型：
--
作者：
Aladag, Nilay;Ozkan-Ariksoysal, Dilsat;Ozsoz, Mehmet
使用7-二甲基氨基-1，2-二苯并吩恶嗪鎓盐（Meldola's blue，MDB）成功地进行了与维生素D受体基因中的阿帕I单核苷酸多态性（SNP）相关的核酸碱基错配的电化学检测，检测限为10.9 pmo 1/100 μ L。探针、错配探针（探针-含SNP的靶）和杂交探针（探针-靶）修饰铅笔石墨电极（PGE）获得的MDB还原信号分别增加。该传感器能够在30分钟的检测时间内清楚地区分完全匹配和错配DNA。研究了影响杂交和指示剂响应的几个因素，以最大限度地提高灵敏度和选择性。与以前的电化学检测DNA杂交的生物传感器的优点进行了讨论。
Electrochemical detection of nucleic acid base mismatches related to Apa I single nucleotide polymorphism (SNP) in the vitamin D receptor gene was performed successfully using 7-dimethyl-amino-1,2-benzophenoxazinium salt (Meldola's blue, MDB) with 10.9 pmo1/100 mu L of detection limit. MDB reduction signals obtained from probe, mismatch(probe-SNP containing target) and hybrid(probe-target) modified pencil graphite electrode(PGE) increased respectively. The sensor was able to clearly distinguish perfect match from mismatch DNA in a 30 min. detection time. Several factors affecting on the hybridization and indicator response are studied to maximize sensitivity and selectivity. The advantages of the biosensor are discussed in comparison with previous electrochemical assays for DNA hybridization.