Voltammetric determination of the Alzheimer’s disease-related ApoE 4 gene from unamplified genomic DNA extracts by ferrocene-capped gold nanoparticles
Voltammetric determination of the Alzheimer’s disease-related ApoE 4 gene from unamplified genomic DNA extracts by ferrocene-capped gold nanoparticles
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用二茂铁封端的金纳米颗粒伏安法测定未扩增基因组 DNA 提取物中与阿尔茨海默病相关的 ApoE 4 基因
DOI：
10.1007/s00604-018-3087-9
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发表时间：
2018
影响因子：
5.7
通讯作者：
Nan Wang
中科院分区：
文献类型：
--
作者：
Hanwen Lu;Ling Wu;Jingrui Wang;Zixiao Wang;Xinyao Yi;Jianxiu Wang;Nan Wang
本文介绍了一种灵敏的检测载脂蛋白E4基因的方法，该方法对阿尔茨海默病的早期诊断具有重要意义。它是基于信号放大，通过使用二茂铁（Fc）封端的金纳米粒子与链霉亲和素修饰。固定化寡核苷酸探针捕获互补apoE 4基因。随后特异性识别被限制酶HhaI水解的GCGC序列。切割仅发生在互补的apoE 4双链体上，而错配阻止酶促切割.因此，apoE 4序列可以与其它apoE序列区分开。得益于Fc加帽纳米颗粒/链霉亲和素的信号放大和HhaI的识别，检测限低至ApoE 4基因的0.1 pM。从血液中提取的四个基因组DNA样品进行了分析的apoE 4基因的存在。这里提出的方法将提供可行的原理证明酶辅助电化学检测基因组DNAs.Graphical abstractionSchematic presentation of amplified voltummetric detection of Alzheimer's disease-related apoE 4 gene from unamplified genome DNA extracts via ferrocene capped gold nanoparticle/streptavidin.
A sensitive method is described for detection of the apoE 4 gene detection which is important for early diagnosis of Alzheimer’s disease. It is based on signal amplification by using ferrocene (Fc) capped gold nanoparticles modified with streptavidin. The immobilized oligonucleotide probe captures complementary apoE 4 gene. This is followed by the specific recognition of the GCGC sequences which are hydrolyzed by the restriction enzyme HhaI. Cleavage only occurs at the complementary apoE 4 duplex, while mismatches prevent enzymatic cleavage. Thus, the apoE 4 sequence can be discriminated against other apoE sequences. Benefitting from amplified signal by Fc-capped nanoparticle/streptavidin and the recognition of HhaI, the detection limit is as low as 0.1 pM of the ApoE 4 gene. Four genomic DNA samples extracted from blood were analyzed for the presence of the apoE 4 gene. The approach presented here will provide viable proof-of-principle for an enzyme-assisted electrochemical assay for the apoE 4 gene in genomic DNAs.Graphical abstractSchematic presentation of amplified voltammetric detection of Alzheimer’s Disease-related apoE 4 gene from unamplified genomic DNA extracts via ferrocene capped gold nanoparticle/streptavidin.