2016-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/659308摘要：對比率懸殊的DNA 混合物中之微量者的鑑別，是法醫基因體學最具挑戰性的議題之一。新發展的巨量平行定序(MPS)能分析混合DNA 檢體，目前應用巨量平行定序(MPS)作人別鑑定的套組已結合常用的STR 及SNP 點位，但是作比率懸殊混合DNA 之微量者分析時，往往得到的微量者特殊點位型別數目不足，無法作微量者的人別鑑定，必須設法增加檢測之STR及SNP 點位數目，尤其是型別定義清楚且能在短片段上檢測的SNP 點位，增加敏感度，才適用於法醫混合DNA 檢體中微量者人別鑑定。本研究以前期研究完成的應用MPS 分析的1214 個SNP 點位組合及過去研發的STR 套組為基礎，目標是發展新的適用於MPS 的多重STR 及SNP 點位組合，能分析比率懸殊混合之DNA 檢體，檢驗其中微量者的特殊型別，且其型別的數量要足以作人別鑑定。我們已收集30 位男性及30 位女性自願捐出者的DNA 檢體，其中半數有近親關係，第一年我們將設計結合約30 個STR 及300 個SNP 點位的適用於MPS 的套組，作約90 份無血緣關係者比率懸殊的混合DNA 檢體型別分析，分辨多量者及微量者的DNA 型別，建立統計模式且計算微量者特殊型別的綜合隨機符合率(RMP)作人別鑑定，並分析其中STR 及SNP 點位的有效性，第二年我們擬調整第一年的STR 及SNP 套組，作約90 份有血緣關係者比率懸殊的混合DNA 檢體及約5 份法醫混合DNA 檢體型別分析，計算微量者特殊型別的綜合RMP作人別鑑定及血緣鑑定。此研究結果將建立新的應用巨量平行定序方法的多重STR 及SNP 點位人別鑑定套組，且建立統計模式增進對比率懸殊混合的親屬及非親屬DNA檢體中微量DNA檢體的基因型別的判別能力，此研究結果在法醫實務應用極為重要。<br> Abstract: To identify the minor component of DNA mixture with very uneven proportion is an importantissue in forensic genomics. Massively parallel sequencing (MPS) can be used for detection of DNAmixture. Presently, there is a combined short tandem repeat (STR) and single nucleotidepolymorphism (SNP) loci individual identification panel for MPS. This panel is effective forindividual identification of a single person. However, the informative polymorphic loci are notsufficient for individual identification of the minor component of DNA mixture with very unevenproportion. In order to obtain sufficient informative polymorphic loci for individual identificationof the minor component, we have to increase the STR and SNP loci detected to increase thesensitivity. Particularly, more SNP loci that have clear information and can be analyzed with shortDNA fragment are necessary.This research is based on our previously established 1214 SNP loci panel using MPS analysis andpreviously established STR panels. We aim to develop a new multiple STR and SNP loci individualidentification panel using MPS. This new panel can be used to detect the genotypes of minorcomponent of DNA mixture with very uneven proportion. The informative genotypes will besufficient for individual identification.We collected 30 DNA samples of male donors and 30 DNA samples of female donors. About halfof these donors are relatives. In the first year, we will establish a panel with 30 STRs and 300 SNPsfor MPS analysis. Ninety DNA mixture samples with very uneven proportion of unrelatedindividuals will be analyzed. The genotypes of alleles of major and minor components will bedetected. The combined RMP (random match probability) of the minor donor will be calculated.The effectiveness of each STR and SNP will be evaluated. In the second year, the STR and SNPloci of the panel will be modified. Ninety DNA mixture samples with very uneven proportion ofrelatives and five forensic DNA mixture samples will be analyzed using MPS. The combined RMPof the minor donor will be calculated based on the genotypes of the minor component detected. Thekinship analysis will be performed for major and minor component based on their relativeness.A new multiple STRs and SNPs individual identification panel using MPS will be established inthis study. This panel will be effective to detect the genotypes of the minor component of DNAmixture with very uneven proportion of relatives and non-relatives. This panel will be very helpfulfor forensic casework.