Yao Juan, Zhang Zhang, Huang Xiaoling, Guo Yongcan
Department of Laboratory Medicine, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou, PR China.
Key Laboratory of Laboratory Medical Diagnostics of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, PR China.
Biosens Bioelectron. 2022 Jul 1;207:114138. doi: 10.1016/j.bios.2022.114138. Epub 2022 Mar 18.
Single nucleotide variants in CYP2C192 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 CYP2C192 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.
CYP2C192中的单核苷酸变异与冠心病患者对氯吡格雷的抵抗有关。为指导药物剂量和个性化用药，首先采用阻断剂置换扩增法特异性扩增G位点和A位点等位基因。对于电化学策略，正向引物标记有电化学活性的亚甲蓝和二茂铁，G位点在-0.26 V产生信号，A位点在0.22 V产生信号。对于侧向流动试纸条检测，使用经过特异性修饰的引物，在测试线1针对G位点产生独特颜色，在测试线2针对A位点产生独特颜色。总之，我们开发了一种基于丝网印刷碳电极的灵敏电化学传感器和基于金纳米颗粒的侧向流动试纸条检测策略，成功对CYP2C192的GG、GA和AA基因型进行基因分型。所提出的方法可实现对包括全血、口腔拭子、唾液和发根在内的多种生物样本的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可视试纸条检测，该平台在资源有限地区的基因分型方面显示出巨大潜力。
