2017-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/705009摘要：本研究將開發新的Y 染色體之多型性STR 基因，提升性侵害案件證物中男性嫌犯人數與人別鑑定之準確度。第一年將尋找新的多型Y-STR 基因，從GeneBank 中四個版本之Y 染色體基因體序列NC_000024.10、NC_000024.9、AC_000156.1 與AC_000067.1 各約有5 千多萬鹼基中尋找，將交叉比對其序列，找出具有高變異之多型性STR 基因，排除上述現行27 個基因，篩選出新的多型性STR 序列，設計PCR 引子複製片段，測試單獨PCR 複製效果、進行複合式PCR 複製、以次世代定序技術（NextGeneration Sequencing, NGS）進行定序測試複合式PCR 複製效果、測試200 個男性DNA 檢體評估多型性並建立對偶基因型序列、篩檢出至少20 組高變異性之Y-STR 基因。第二年將針對現行27 組Y-STR基因設計適合NGS 系統使用之PCR 複製引子，測試PCR 複製效果、測試複合式PCR 複製效果、將新開發之20 組合併成47 組Y-STR 基因系統、以NGS 進行定序、建立NGS 之Y-STR 基因數據分析系統、測試男女性DNA 混合型之NGS 系統檢測靈敏度、測試多名男性DNA 混合型之NGS 系統檢測靈敏度與分辨能力、建立NGS 系統之確效試驗數據等。預估本47 組Y-STR 基因系統之隨機相符頻率將在10 至100 萬分之一間，可大幅提升性侵嫌犯人數與人別鑑定之準確度，且以NGS 鑑定亦可提升鑑定靈敏度與篩檢大量檢體之效率。<br> Abstract: This study will develop novel highly polymorphic Y-STR loci to increase the reliability of estimating thenumber of involved males and their personal identification. The work of the first year is to find novelpolymorphic Y-STR loci. The novel polymorphic STR loci will be sorted by the comparison of 4 genomesequences of Y chromosome from GeneBank (NC_000024.10, NC_000024.9, AC_000156.1 andAC_000067.1) around 50 million bases of each. The abovementioned 27 Y-STR loci will be excluded. Theoptima of primer sequences and PCR condition will be tested. Multiplex PCR will be tested for evaluationthe interaction of primers among loci. Next Generation Sequencing (NGS) will be used to evaluate results ofmultiplex PCR. Two hundred male DNA will be screened to evaluate the polymorphism of STR and establishallele sequences. At least 20 highly polymorphic Y-STR loci will be established. In the second year, the novel20 Y-STR loci will combine currently used 27 Y-STR loci in a 47-plex Y-STR genotyping system. Primers of27 Y-STR loci for small size amplicons will be designed for NGS system. Single PCR will be tested forevaluation of primer. NGS will be used to evaluate results of multiplex PCR. Optima of primer sequences orprimer concentration of 47 Y-STR loci will be tested to establish a robust typing system. Data analysis modelwill be established. Sensitivity of different mixed ratio of male and female DNA and discrimination power ofmixture with multiple male DNA will be performed. Developmental validation will be studied for forensicuse. According to the data of 27 Y-STR loci system, the random match probability of the 47-plex Y-STRgenotyping system will be around one in 0.1 million to 1 million. It will greatly increase the reliability ofsuspect identification in sexual assault cases. Besides, the sensitivity of detection and efficiency of massscreening will be also greatly increased using NGS technology.