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

色盲是一种x连锁遗传病，影响约5- 10%的男性。据估计，女性携带者约占日本人口的10%。基因分析是在没有家谱信息的情况下识别女性携带者的唯一方法。本研究的目的是建立一个可靠的方案，以确定女性携带者。从志愿者口腔黏膜拭子中提取DNA样本。采用改进的组织基因组DNA提取方案提取基因组DNA。阳性对照PCR显示-肌动蛋白基因在局部粘膜拭子样本中扩增成功。设计了视蛋白基因引物，用于PCR分析。虽然获得了几个PCR产物，但DNA测序结果显示这些产物均为非特异性扩增。这一结果表明，引物设计需要进一步改进才能有效扩增。从口腔粘膜拭子样本中成功扩增出其他几个基因，因此我们的样本收集和DNA提取方案被认为是充分和可靠的诊断使用。

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.