Establishment of genetic diagnosis protocol for color blindness using oral mucosa scratch sample

口腔粘膜划痕样本色盲基因诊断方案的建立

• 批准号: 15591926
• 负责人: SAKAMOTO Kei
• 金额: $ 2.24万
• 依托单位: Tokyo Medical & Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591926/
• 关键词: Color blindness; Buccal mucosa swab; Gene diagnosis; 口腔粘膜擦過細胞; 遺伝子診断法; オプシン

Color blind is an X-linked genetical disease that affects approximately 5-10 % males. Female carrier is estimated to count approximately 10 % of the Japanese population. Gene analysis is the only way to identify a female carrier when family pedigree information is not available. The purpose of the present study is to establish a reliable protocol to identify a female carrier. DNA sample was obtained from buccal mucosa swab of volunteers. Genomic DNA was extracted using modified protocol for tissue genomic DNA extraction. Positive control PCR for beta-actin gene revealed successful amplification from buocal mucosa swab samples. Primers for opsin gene was designed and used for PCR analysis. Although several PCR products were obtained, DNA sequencing results showed that these products were non-specific amplification. This result indicate sthat further refinement of primer design is require for efficient amplification. Several other genes were successfully amplified from buccal mucosa swab samples, thus our sample collection and DNA extraction protocol was thought to be sufficient and reliable for diagnostic use.

色盲是一种X连锁遗传疾病，影响大约5- 10%的男性。女性携带者估计约占日本人口的10%。基因分析是唯一的方法来确定一个女性携带者时，家庭系谱信息不可用。本研究的目的是建立一个可靠的协议，以确定女性携带者。从志愿者的颊粘膜拭子中获得DNA样品。使用改良的组织基因组DNA提取方案提取基因组DNA。β-肌动蛋白基因的阳性对照PCR显示从粘液粘膜拭子样品中成功扩增。设计了视蛋白基因的特异性引物，并进行了PCR分析。虽然获得了多个PCR产物，但DNA测序结果显示这些产物为非特异性扩增。这一结果表明，为了有效扩增，需要进一步改进引物设计。从颊粘膜拭子样本中成功扩增了几个其他基因，因此我们的样本收集和DNA提取方案被认为是诊断用途的充分和可靠的。