Department of Bio-analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Japan.
Ann Clin Biochem. 2014 Mar;51(Pt 2):298-300. doi: 10.1177/0004563213499316. Epub 2013 Sep 18.
The proportion of poor metabolizers (PMs) of cytochrome P450 (CYP) 2C19 is much higher in the Japanese population than in European populations. Cycling probe technology (CPT) is a simple signal amplification technique for targeting specific DNA sequences. CPT utilizes a chimeric DNA-RNA-DNA probe that is cleaved by the enzyme ribonuclease (RNase H). In this study, using CPT, we aimed to detect the CYP2C19 gene polymorphism from noninvasive samples to determine extensive metabolizers (EMs) and PMs of CYP2C19.
DNA samples were extracted from hair, buccal mucosa, and blood cells. Primers and cycling probes were designed specifically for region G636A for exon 4 and G681A for exon 5, reported to be gene polymorphisms of CYP2C19.
DNA extracted from hair follicle cells and buccal epithelial cells was the same as that collected from invasive blood sampling. The genotype of CYP2C19 was successfully identified as either EM or PM in 71 samples, producing identical results to those for the TaqMan method, except in three samples.
We successfully detected the two gene polymorphisms of CYP2C19 from noninvasive samples using a simple DNA extraction method and CPT.
细胞色素 P450（CYP）2C19 的代谢不良者（PM）在日本人群中的比例远高于欧洲人群。循环探针技术（CPT）是一种针对特定 DNA 序列的简单信号放大技术。CPT 利用一种嵌合 DNA-RNA-DNA 探针，该探针可被核糖核酸酶（RNase H）切割。在这项研究中，我们使用 CPT 从非侵入性样本中检测 CYP2C19 基因多态性，以确定 CYP2C19 的广泛代谢者（EM）和 PM。
从头发、口腔黏膜和血细胞中提取 DNA 样本。针对exon 4 的 G636A 和 exon 5 的 G681A 报道为 CYP2C19 基因多态性，设计了专门的引物和循环探针。
从毛囊细胞和口腔上皮细胞中提取的 DNA 与从侵入性血液采样中收集的 DNA 相同。在 71 个样本中，成功地将 CYP2C19 的基因型鉴定为 EM 或 PM，除了三个样本外，其结果与 TaqMan 方法相同。
我们使用简单的 DNA 提取方法和 CPT 从非侵入性样本中成功检测到 CYP2C19 的两个基因多态性。
