2015-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/701776摘要：本研究將發展以次世代定序技術（Next Generation Sequencing, NGS )進行大量STR ( short tandem repeat)基因鑑定之研究，由於NGS分析序列片段（read)之長度已可達到400 bp，可以涵蓋高變異 STR基因對偶基因型的長度，因此NGS在未來極可能取代現行鑑定STR基因長度之毛細管電泳法， 以高通量之NGS同步鑑定大量檢體中之大量STR基因，大幅提高法醫人別鑑定之效能。本研究將以 三年時間建立此系統，第一年將針對NGS分析出之大量fasta與fastq片段序列數據開發出快速尋找 STR序列之軟體系統，並建立STR基因變異分類中10個單一重複單位（simple repeat) STR基因類之 多重PCR複製組合系統，比較毛細管電泳法與NGS法鑑定結果。第二年將建立分析STR基因變異分 類中11個複雜之多種重複單位（complex repeat) STR基因類之多重PCR複製組合系統，與第一年之 10個基因組合為21個多重PCR複製組合系統，並比較毛細管電泳法與NGS法鑑定結果，並發展直 接由NGS分析出之大量fasta與fastq片段序列數據快速辨識STR基因差異係重複序列差異或SNP序 列差異之軟體系統。第三年將建立另外12組複雜之多種重複單位STR基因類之多重PCR複製組合系 統，與第一、二年之21個基因組合為33個多重PCR複製組合系統，建立以NGS法分析STR基因之 模式。並建立可以選擇捜尋序列之比對模式在數百萬個fasta/fastq數據中，快速判定33個STR基因 約500餘對偶基因型之分析軟體。<br> Abstract: This study will develop a system of massive STR genotyping by Next Generation Sequencing (NGS) for forensic applications. Since the high throughput and length of read obtained by NGS has been increased to 400 bp which covers the length of STR loci, it is very likely to replace capillary electrophoresis by NGS technique in STR genotyping in the near future. This will greatly increase the efficiency of DNA identification in forensic applications. This study will take 3 years. The software reading STR features of fasta/fastq generated by NGS will be developed in the first year. Besides, multiplex genotyping of 10 STR loci with simple repeat feature will be established to test the performance of the NGS sequence mapping system. The accuracy between capillary electrophoresis and NGS techniques will be performed. The study of the second year will develop the multiplex genotyping of another 11 STR loci with complex repeat feature and combine with 10 multiplex STR loci developed in the first year. The alleles with non-consensus repeat will be discovered and recorded by the NGS sequence mapping system. In the third year, this study will develop the multiplex genotyping of another 12 STR loci with complex repeat feature and generate a STR genotyping system with 33 loci which covers the most popular STR loci used in forensic laboratories. The NGS sequence mapping system will be mature enough to identify more than 500 alleles of 33 loci in millions fasts/fastq data of NGS.次世代定序短重複序列人別鑑定法醫鑑定next generation sequencingshort tandem repeatpersonal identificationforensic medicine