School of Pharmaceutical Sciences, Mukogawa Women's University.
Institute of Biosciences, Mukogawa Women's University.
Biol Pharm Bull. 2019;42(8):1345-1349. doi: 10.1248/bpb.b19-00110.
DNA extraction and purification have been generally considered to be required for PCR assay. We demonstrated a new protocol using biological specimens directly as templates for real-time PCR with melting curve analysis. We confirmed the melting curve analysis was particularly suitable for the identification of the insertion/deletion (Ins/Del) polymorphism of the angiotensin-converting enzyme (ACE) gene. The new protocol we developed can be set up using simple and complete PCR analysis including data interpretation in under four hours with additional advantages of application for large-scale clinical research, diagnostics, and epidemiological studies at low cost.
DNA 提取和纯化通常被认为是 PCR 检测所必需的。我们展示了一种新的方案，该方案使用生物样本直接作为模板进行实时 PCR 及熔解曲线分析。我们证实熔解曲线分析特别适合血管紧张素转换酶（ACE）基因插入/缺失（Ins/Del）多态性的鉴定。我们开发的新方案可以在不到四个小时的时间内完成，包括简单而完整的 PCR 分析和数据解释，具有应用于大规模临床研究、诊断和低成本流行病学研究的额外优势。
