Faculty of Pharmacy, Analytical Chemistry Department, Ege University, Bornova, Izmir 35100, Turkey; Biotechnology Department, Graduate School of Natural and Applied Sciences, Ege University, Bornova, Izmir 35100, Turkey.
Faculty of Pharmacy, Analytical Chemistry Department, Ege University, Bornova, Izmir 35100, Turkey; Biotechnology Department, Graduate School of Natural and Applied Sciences, Ege University, Bornova, Izmir 35100, Turkey.
Int J Biol Macromol. 2019 Mar 15;125:839-846. doi: 10.1016/j.ijbiomac.2018.12.107. Epub 2018 Dec 13.
Single nucleotide polymorphisms (SNPs) are the most common type of genetic variation among people. Development of reliable methods for the detection of SNP is crucial in aspects of molecular diagnosis and personalized medicine. In our study, a genomagnetic assay in combination with zip nucleic acid (ZNA) for electrochemical detection of SNP related to Factor V Leiden mutation. For the first time in the literature, a new generation nucleic acid; ZNA was applied herein for electrochemical monitoring of nucleic acid hybridization. Streptavidin coated magnetic beads (MBs) were used for preparation of samples containing ZNA-DNA hybrid and accordingly, the guanine signal was measured as a response of hybridization related to Factor V Leiden mutation by carbon nanofibers (CNF) modified screen printed electrodes (SPE) and multi-channel screen printed array of electrodes (CNF-MULTI SPEx8). The detection limit (DL) was found to be 3.79 μg/mL (376 nM) and, 11.63 μg/mL (1.624 μM), respectively by CNF-SPE and CNF-MULTI SPEx8. The selectivity of ZNA probe to mutation-free DNA sequences was also investigated in contrast to DNA probe. The applicability of ZNA based magnetic beads assay to sequence selective hybridization related to Factor V Leiden was also tested in synthetic PCR samples.
单核苷酸多态性（SNPs）是人与人之间最常见的遗传变异类型。开发可靠的 SNP 检测方法对于分子诊断和个性化医疗方面至关重要。在我们的研究中，结合基因组磁珠法和拉链核酸（ZNA）电化学检测与因子 V Leiden 突变相关的 SNP。本文首次应用新一代核酸；ZNA 用于电化学监测核酸杂交。链霉亲和素包被的磁珠（MBs）用于制备含有 ZNA-DNA 杂交体的样品，并且通过碳纤维纳米纤维（CNF）修饰的丝网印刷电极（SPE）和多通道丝网印刷电极阵列（CNF-MULTI SPEx8）测量与因子 V Leiden 突变相关的杂交的鸟嘌呤信号。通过 CNF-SPE 和 CNF-MULTI SPEx8，检测限（DL）分别为 3.79μg/mL（376nM）和 11.63μg/mL（1.624μM）。还研究了 ZNA 探针与无突变 DNA 序列的选择性，与 DNA 探针相比。还在合成 PCR 样品中测试了基于 ZNA 的磁珠测定法在与因子 V Leiden 相关的序列选择性杂交中的适用性。
