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Toehold-Mediated Strand Displacement Reaction for Dual-Signal Electrochemical Assay of Apolipoprotein E Genotyping.
ACS Sensors ( IF 8.2 ) Pub Date : 2020-09-01 , DOI: 10.1021/acssensors.0c01511
Hanwen Lu 1 , Binrong Ding 2 , Liujuan Tong 1 , Fan Wu 1 , Xinyao Yi 1 , Jianxiu Wang 1
Affiliation  

Apolipoprotein E (apoE) polymorphic genes are one of the main genetic determinants of Alzheimer’s disease (AD) risk. Relying on the toehold-mediated strand displacement reaction (SDR), the dual-signal electrochemical assay of apoE genotyping with potential applications in the early diagnosis of AD has been achieved. The displacement of the surface-confined methylene blue- and ferrocene-capped detection probe-modified gold nanoparticles (AuNPs) by the complementary sequences (Tc 1 and Tc 2, fragment of allele ε4 at codon 112 and that of allele ε3 or ε4 at codon 158, respectively), triggered by the highly specific SDR, results in decreased voltammetric signals. In contrast, partial strand displacement caused by the single mismatched sequences (Tsm 1 and Tsm 2, fragment of allele ε2 or ε3 at codon 112 and that of allele ε2 at codon 158, respectively) produces larger voltammetric signals. The proposed method serves as a versatile platform for the discrimination of six apoE genotypes, including three homozygotes (ε2/2, ε3/3, and ε4/4) and three heterozygotes (ε2/3, ε2/4, and ε3/4), and for the quantification of apoE ε3/3 from genomic DNA extracts of AD patients.

中文翻译:

脚趾介导的链置换反应用于载脂蛋白E基因分型的双信号电化学分析。

载脂蛋白E(apoE)多态性基因是阿尔茨海默氏病(AD)风险的主要遗传决定因素之一。依靠脚趾介导的链置换反应(SDR),实现了apoE基因分型的双信号电化学分析,并有望在AD的早期诊断中应用。表面限制的亚甲基蓝和二茂铁加帽的检测探针修饰的金纳米颗粒(AuNPs)的互补序列(Tc 1和Tc 2,等位基因ε4的第112位密码子片段和等位基因ε3或ε4的密码子)分别由高特异性SDR触发158)导致伏安信号降低。相反,部分链位移是由单个错配序列(Tsm 1和Tsm 2,等位基因ε2或ε3的第112位密码子和等位基因ε2的密码子158,分别产生较大的伏安信号。所提出的方法为区分六个apoE基因型提供了一个通用平台,包括三个纯合子(ε2/ 2,ε3/ 3和ε4/ 4)和三个杂合子(ε2/ 3,ε2/ 4和ε3/ 4)。 ,并用于从AD患者的基因组DNA提取物中定量apoEε3/ 3。
更新日期:2020-09-25
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