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

2004 Fiscal Year Final Research Report Summary

• Project/Area Number: 15591926
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Morphological basic dentistry
• Research Institution: Tokyo Medical & Dental University
• Principal Investigator: SAKAMOTO Kei Tokyo Medical & Dental University, Research assistant, 大学院・医歯学総合研究科, 助手 (00302886)
• Co-Investigator(Kenkyū-buntansha): KATSUBE Ken-ichi Tokyo Medical & Dental University, Lecturer, 大学院・医歯学総合研究科, 講師 (20233760)
• Project Period (FY): 2003 – 2004
• Keywords: Color blindness / Buccal mucosa swab / Gene diagnosis

Research Abstract

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.