2012-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/705220摘要：在法醫學領域中，應用各種DNA 多型性標記作人身鑑別及親屬血緣鑑定是重要的發展主題，目前常規性應用作人身鑑別及血緣鑑定的方法包括體染色體及Y染色體短多重複序列標記及粒線體DNA非轉譯區(多變異I 區及多變異II 區)的定序。因為這些標記或DNA 序列在不同的族群都呈現豐富的多型性，所以能提供大量的多型性資訊作鑑定判斷的基礎。但是，短多重複序列標記較易突變，較難合併在一個PCR 反應中完成，而需要較多量的檢體及較長DNA 片段(200~500bp)，但是法醫學個案的DNA 檢體，常常是微量及被破壞而斷裂的，所以這些常規性的方法應用於法醫學科案件檢驗有時得不到完整結果而無法鑑定。近年來，單一核苷酸多型性(SNP)被發現可應用於人身鑑別，應用單一核苷多型性的優點包括突變率低，只需較短的DNA 片段(少於150 核苷酸序列)即可判斷，所以，法醫學的案件中斷裂的DNA片段也可以適用，而且短片段的DNA，可以應用高產出能力的基因分析技術，目前許多研究在發展新的體染色體、性染色體及粒線體DNA SNP 位置組合，用於人身鑑別及親緣鑑定，而SNP 多型性在不同族群可能不同，如何挑選出在不同族群都有高變異性比例的SNP 點位，組合成高鑑別力的SNP分析系統是重要的研究主題。我們的研究目的為建立新的粒線體、體染色體及性染色體各有50~70 個高變異性的SNP 標記array晶片的組合，且總應用於不同族群的人身鑑別及親屬鑑定。我們的研究材料包括12 個法醫學常用的CEPH 細胞株及1000 個在台灣生活、無血緣關係自願者捐出的DNA 檢體。我們研究的第一年，要收集DNA 檢體及篩檢現存資料庫的高度變異比例的SNP點位，選出50~70 體染色體DNA SNP 點位，作成一組array 晶片，在我們的1000 個DNA 檢體及12cell-line DNA 檢體分析，最後試用於100 個真正法醫DNA 斷裂案件檢體，檢測其適用性。第二年，篩檢現存資料庫的高度變異比例的SNP 點位，選出50~70 性染色體DNA SNP 點位，作成一組array晶片，在我們的1000 個DNA 檢體及12 cell-line DNA 檢體分析，最後試用於100 個真正法醫DNA 斷裂案件檢體，檢測其適用性。第三年，經過篩檢現存資料庫的高度變異比例的SNP 點位，作成一組array 晶片，選出的50~70 粒線體DNA SNP 點位，在我們的1000 個DNA 檢體及12 cell-line DNA 檢體分析，最後試用於100 個真正法醫DNA 斷裂案件檢體，檢測其適用性。我們還要把結果建立體染色體、性染色體、粒線體之SNP 點位型別資料庫。研究結果將作成報告及發表在國際期刊，我們預期將能建立有變異性高、高鑑別力及高親子指數的粒線體、體染色體及性染色體SNP 標記組合，可應用於人身鑑別及親緣鑑定，且在不同族群皆可應用。<br> Abstract: Genetic tests with polymorphic DNA makers of human identification are critical to the field of forensicscience and parentage or relativeness testing . Autosomal and Y chromosomal short tandem repeat (STR)makers and mitochondrial non-coding region (HV I AND HV II) sequences are routinely used currently,These markers are highly informative because of the large number of alleles within various populations andthe established database of different population groups. However, STR markers are limited by their highmutation rate, difficulty with regards to multiplexing, and the need for large amplification products, whichlimits the use of degraded samples in forensic cases.Single nucleotide polymorphisms (SNPs) have been promoted as useful genetic markers for humanidentification recently. Single nucleotide polymorphisms have low mutation rates and rely on shortamplicons (i.e., less than 150bp), which allow for the use of degraded DNA samples in forensic case workand high-throughput genotyping technologies. Several studies have described preliminary SNP panels,containing autosomal, sex chromosomal or mitochondrial SNP markers, for use in human identification andparentage testing. Furthermore, selecting a SNP with high heterozygosity and low differences in allelefrequency for an individual population group or for all population is important for the development of anefficient SNP marker system with high discrimination power.The aim of our study is to establish sets of mitochondrial, autosomal, and sex chromosomal SNP markersarray system with 50~70 high heterozygosity for human identification and parentage testing in differentpopulation groups.We will collect DNA samples of 12 cell-lines from CEPH families and 1000 DNA samples from theunrelated volunteer donors living in Taiwan.In the first year, DNA samples will be collected. A set of array with fifty to seventy candidate autosomalSNP loci will be established and analyzed in our 1000 DNA samples and DNA samples of cell lines.Validation will be performed in 100 forensic DNA samples with degraded DNA. For the second year, a setof array with 50-70 candidate sex-chromosomal SNP loci will be established and analyzed in our 1000 DNAsamples and DNA samples of cell lines. Validation will be performed in 100 forensic DNA samples withdegraded DNA. In the third year, a set of array with 50-70 candidate SNP loci of mitochondrial DNA will beestablished and analyzed in our 1000 DNA samples and DNA samples of cell lines. Then, validation will beperformed in 100 forensic DNA samples with degraded DNA. Database of these mitochondrial, autosomaland sex-chromosomal SNP markers will be established.The results will be reported. Our findings will present that highly polymorphic SNP markers have a highcombined power of identity (CPI) value and high discrimination power for parentage testing and humanidentification, respectively. In addition, highly informative SNP markers with high heterogeneity will beproved to be sufficient for human identification in diverse human populations.單一核苷酸多型性人身鑑別晶片分析single nucleotide polymorphismhuman identificationarray