Wang Haobo, Yan Chi, Tong Hua, Ye Xiaojun, Yuan Xiao, Liu Cui, Li Hongbo
School of Materials Science and Engineering, East China University of Science and Technology Shanghai 200237 China
RSC Adv. 2024 Aug 19;14(36):26007-26015. doi: 10.1039/d4ra03913k. eCollection 2024 Aug 16.
The rapid detection of single nucleotide polymorphisms (SNPs) in the CYP2C19 gene is crucial for precise clopidogrel usage. Quantitative real-time polymerase chain reaction (qPCR), as a powerful amplification tool, has been widely employed for CYP2C19 SNPs detection. However, traditional qPCR suffers from long amplification times and high reagent consumption. To address these challenges, this work presents a microfluidic SNPs detection device based on on-chip qPCR. The device includes a rapid thermal cycling system, an optical detection system, a control system, and a complementary silicon-glass chip for CYP2C19 SNPs detection. Compared to commercial qPCR instruments that take 1 hour for testing, this device completes the test in just 15 minutes (40 PCR cycles). The resulting linearity is similar to that found using commercial qPCR instruments but with higher amplification efficiency. Additionally, compared with other silicon-based qPCR chips, this chip is constructed by using a convenient two-step method and offers low manufacturing costs, which potentially reduces single-test costs to an acceptable level. This makes our chip promising for point-of-care testing (POCT).
快速检测细胞色素P450 2C19（CYP2C19）基因中的单核苷酸多态性（SNP）对于精准使用氯吡格雷至关重要。定量实时聚合酶链反应（qPCR）作为一种强大的扩增工具，已被广泛用于检测CYP2C19基因的SNP。然而，传统的qPCR存在扩增时间长和试剂消耗量大的问题。为应对这些挑战，本研究提出了一种基于芯片上qPCR的微流控SNP检测装置。该装置包括一个快速热循环系统、一个光学检测系统、一个控制系统以及用于检测CYP2C19基因SNP的互补硅玻璃芯片。与需要1小时进行检测的商用qPCR仪器相比，该装置仅需15分钟（40个PCR循环）即可完成检测。其线性结果与商用qPCR仪器相似，但扩增效率更高。此外，与其他基于硅的qPCR芯片相比，该芯片采用便捷的两步法构建，制造成本低，这有可能将单次检测成本降低到可接受的水平。这使得我们的芯片在即时检测（POCT）方面具有广阔前景。
