焦磷酸测序技术检测短片段牙釉质蛋白基因进行性别鉴定的分析-法医学专业论文.docxVIP

英 英 文 摘 要 PAGE 4 PAGE 4 Short fragment of amelogenin gene tested by pyrosequencing for gender identification ABSTRACT Objective: Gender information is one of the most important information in the forensic casework and archaeological research. Usually if the corpse is unbroken, forensic workers can make a determination by morphological analysis. However, in some disasters of large group, when the integrity of the human skeleton is destroyed, or bone hypoplasia as a child le ad to identification by morphological analysis difficulty, using molecular biological methods to indentify gender becomes particularly important. At present, the commonly used DNA techniques in gender identification include Y chromosome-specific probes, Zin-finger protein gene (ZFY/ZFX) and amelogenin gene test, among which amelogenin test is the most widely applied. Since 1991 amelogenin gene had been sequenced by Nakahori, Nakahori (1991), Akane (1991), Bailey (1992), Sullivan (1993) had used PCR method to amplify the single copy X-Y homologous regions of amelogenin gene by different primers in sex determination. The method made by Sullivan with PCR products being 106bp and 112bp, has been widely used for gender identification, which was applied by most of the forensic personal identification kits as a sex identification method. However, these existing technologies can not fully meet the needs of a number of difficult cases, and usually can not get clear typing results for highly-degraded samples or ancient DNA, or can lead to invalid amplification result from amelogenin sequence mutation and sex chromosome variation. To solve these problems, this study established a new gender DNA analysis methods, namely, a 45bp stretch of amelogenin gene was chosen for analysis by pyrosequencing. At the same time, a series of validation experiments, such as the sensitivity, accuracy and species specificity, degradation models, bone samples, were performed for the novel method, with a view to provide a ref