Blocker displacement amplification mediated PCR based screen-printed carbon electrode biosensor and lateral flow strip strategy for CYP2C192 genotyping
Blocker displacement amplification mediated PCR based screen-printed carbon electrode biosensor and lateral flow strip strategy for CYP2C192 genotyping
复制标题
基于阻断剂置换扩增介导的 PCR 的丝网印刷碳电极生物传感器和侧流条策略用于 CYP2C192 基因分型
DOI：
10.1016/j.bios.2022.114138
复制
发表时间：
2022
影响因子：
12.6
通讯作者：
郭永灿
中科院分区：
文献类型：
--
作者：
姚娟;张章;黄晓玲;郭永灿
CYP2C19*2 中的单核苷酸变异与冠心病中的氯吡格雷耐药相关。为了指导药物剂量和个体化用药，首先使用阻断剂置换扩增来特异性扩增G位点和A位点等位基因。对于电化学策略，正向引物被标记为电化学活性甲基蓝和二茂铁，在 G 位点的 -0.26 和 A 位点的 0.22 V 上产生信号。对于侧流试纸条测定，使用具有特定修饰的引物在G位点的测试线1和A位点的测试线2中产生独特的颜色。总之，我们开发了一种基于灵敏丝网印刷碳电极的电化学传感器和基于金纳米颗粒的侧流条检测策略，成功地对 CYP2C19*2 GG、GA 和 AA 基因型进行了基因分型。该方法可实现全血、颊拭子、唾液、发根等多种生物样本的CYP2C19*2分析，且与测序结果具有良好的一致性。由于我们提出的策略仅依赖于热循环仪和视觉条检测，因此该平台在源有限区域基因分型中显示出巨大的潜力。
Single nucleotide variants in CYP2C19*2 are associated with clopidogrel resistance in coronary heart disease. In order the guidance the dosage of drug and personalized medicine, blocker displacement amplification was first used to specific amplify G site and A site alleles. For electrochemical strategy, forward primers were labeled electrochemical active methyl blue and ferrocene, generates signals on −0.26 for G site and 0.22 V for A site. For lateral flow strip assay, primers with specific modification were used to generates unique color in test line 1 for G site and test line 2 for A site. In conclusion, we developed a sensitive screen-printed carbon electrodes based electrochemical sensor and gold nano particle based lateral flow strip assay strategy to successfully genotyping CYP2C19*2 GG, GA and AA genotype. The proposed method can realize CYP2C19*2 analysis from multiple biological samples including whole blood, buccal swab, saliva and hair root, and showed good consistency with Sequencing. Due to the fact our proposed strategy merely relies on thermal cycler instrument and visual strip detection, this platform shows great potential in source-limited regions genotyping.