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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 華筱玲(Hsiao-Lin Hwa) | |
dc.contributor.author | Yu-Hsuan Hsieh | en |
dc.contributor.author | 謝宇宣 | zh_TW |
dc.date.accessioned | 2021-06-16T08:17:14Z | - |
dc.date.available | 2020-08-27 | |
dc.date.copyright | 2020-08-27 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-07-13 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58494 | - |
dc.description.abstract | 法醫分子生物技術是人別鑑定與親緣鑑定主要方法,目前是以毛細管電泳為基礎的短相連重複序列 (STR) 分析為主,然而DNA因腐敗或被破壞而裂解時,用傳統毛細管電泳方法所做出的STR基因座分析,結果可能不足以作人別鑑定。近年因巨量平行定序方法 (MPS) 的發展,可以同時分析STR與單核苷酸多型性 (SNP),除了可以得到法醫常規分析的STR基因座資訊之外,還可以將其重複序列進行定序以尋找其序列之變異性,對於腐敗或被破壞而裂解的DNA檢體分析是具有潛力的方法。本研究的目的在透過次氯酸鈉裂解DNA檢體或模擬法醫DNA檢體的分析,探討巨量平行定序方法分析裂解的DNA檢體作人別鑑定及親緣關係鑑定之正確性。 本研究收集來自21組家庭共50位台灣漢人的周邊血、臍帶血、病理切片、舊臍帶及牙齒檢體共81件,其中將部分的周邊血製作成血片並以次氯酸鈉浸泡破壞,而牙齒檢體則直接抽取DNA或是製成脫鈣的牙齒切片後再抽取DNA。分別進行傳統毛細管電泳方法與巨量平行定序方法分析,使裂解與未裂解參考檢體比對,進行人別鑑定及親緣鑑定計算。 研究結果顯示巨量平行定序分析27個體染色體STR、25個Y染色體STR、7個X染色體STR與96個體染色體SNP,其在未裂解參考檢體的分析上皆較毛細管電泳15個體染色體STR分析有更佳的鑑別力。經次氯酸鈉裂解的DNA檢體,雖然在巨量平行定序法上出現較多的對偶基因型改變,然而其在人別鑑定及親緣鑑定計算可以得到更低的隨機相符頻率及更高的累積似然比。而模擬法醫檢體,除了脫鈣的牙齒切片在本研究幾乎無法進行正確的分析之外,一般病理切片、臍帶、一般牙齒皆可得到巨量平行定序法較毛細管電泳法優異的統計結果。 此外本研究也完成了143個台灣地區漢人 (54個男性,89個女性) 以國際法醫遺傳學協會所建議之標準化命名方式進行命名的巨量平行定序 (MPS) 27個體染色體STR、25個Y染色體STR、7個X染色體STR及96個體染色體SNP之頻率資料庫,可以對我國未來的法醫DNA人別鑑定有所助益。 本研究的結論為巨量平行定序方法可以同時分析STR及SNP標記,分析裂解的DNA檢體,作人別鑑定與親緣鑑定,比毛細管電泳法只作STR標記有更好的鑑別效果。 | zh_TW |
dc.description.abstract | Forensic molecular biotechnology is the main method for individual identification and relationship test. It is mainly analysis by short tandem repeat (STR) which is based on capillary electrophoresis. However, when DNA is degraded due to decompose or destroyed, the results of STR loci carried out by traditional capillary electrophoresis may be insufficient for individual identification. In recent years, due to the development of massive parallel sequencing (MPS), STR and single nucleotide polymorphism (SNP) can be analyzed at the same time. In addition to the information of STR loci that can be analyzed routinely, MPS can also sequence the repeated sequence to obtain the information of sequence variances. It is a potential method for the analysis of decomposed or destroyed DNA samples. The purpose of this study is to analyze the accuracy of the individual identification and relationship test by analyzing the DNA samples destroyed by sodium hypochlorite or mock forensic DNA samples. In this study, a total of 81 samples were collected from 50 Taiwanese Han people of 21 families. These samples included peripheral blood, umbilical cord blood, pathological sections, dried umbilical cord and dental specimens. Dried blood spots were made and soaked with sodium hypochlorite to destroy DNA. DNA of the dental specimens were extracted directly or extracted after decalcified tooth slices were prepared. The traditional capillary electrophoresis method and MPS method were performed to analyze the degraded and nondegraded samples, and the individual identification and relationship test were also performed. The results show that MPS method with 27 autosomal STRs, 25 Y-STRs, 7 X-STRs and 96 autosomal SNPs analysed has better discrimination in the analysis of nondegraded samples than using capillary electrophoresis method with 15 autosomal STRs analysed. Although the DNA samples destoryed by sodium hypochlorite showed more alleles changed in the MPS method, the individual identification and relationship test still showed better discrimination than capillary electrophoresis method. Except for decalcified tooth slices, which can hardly be accurately analyzed in this study, mock forensic DNA samples included pathological sections, dried umbilical cords, and teeth could obtain excellent statistical results from MPS method compared to capillary electrophoresis method. In addition, the allele frequency database for MPS of 143 random Taiwanese Han people (54 males, 89 females) was also described in this study. The alleles of this database included 27 autosomal STRs, 25 Y-STRs, 7 X-STRs and 96 autosomal SNPs, which were named after the standardized naming method recommended by International Society for Forensic Genetics. The database would be helpful to Taiwan in individual identification. In conclusion, MPS method is able to analyze the degraded DNA samples for individual identification and relationship test. | en |
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dc.description.tableofcontents | 國立臺灣大學碩士學位論文 口試委員會審定書 . I 致謝 .. II 中文摘要 . III Abstract. V 第一章 序言. 1 第二章 研究目的與文獻回顧.. 3 第一節 法醫學人別鑑定(forensic DNA identification). 3 一、 司法分子生物學簡介. 3 二、 基因變異性(genetic variation) 及DNA 的多型性(DNA polymorphism). 3 三、 法醫人別鑑定方法. 4 第二節 巨量平行定序方法(massively parallel sequencing, MPS) . 6 一、 巨量平行定序方法簡介 . 6 二、 標準化命名方法 . 7 三、 族群頻率資料庫. 8 第三節 人為破壞之法醫檢體 . 9 一、 次氯酸鈉. 9 二、 福馬林. 10 第四節 研究目的. 11 第三章 實驗材料與方法. 12 第一節 實驗設計. 12 第二節 實驗材料. 12 第三節 實驗方法. 12 一、 次氯酸鈉浸泡破壞. 12 二、 DNA 萃取. 13 三、 DNA 定量. 15 四、 DNA 基因分析. 17 第四節 資料分析與統計. 19 一、 短相連重複序列(STR) 資料分析. 19 二、 巨量平行定序(MPS) 資料分析. 19 三、 台灣地區漢人巨量平行定序(MPS) 頻率資料庫建立. 20 四、 人別鑑定與親緣鑑定分析. 20 第四章 實驗結果. 22 第一節 檢體破壞、DNA 萃取及定量結果. 22 第二節 以毛細管電泳法分析STR. 23 一、 以毛細管電泳法分析次氯酸鈉裂解DNA 檢體STR 之結果. 24 二、 以毛細管電泳法分析模擬法醫DNA 檢體STR 之結果. 25 第三節 巨量平行定序(MPS) 法之品質評估. 26 第四節 以NGS 分析STR 與SNP. 27 一、 以MPS 分析次氯酸鈉裂解DNA 檢體之STR 及SNP 結果. 27 二、 以MPS 分析模擬法醫DNA 檢體之STR 及SNP 結果. 29 第五節 台灣地區漢人巨量平行定序(MPS) 頻率資料庫. 32 一、 以ForenSeq™ DNA Signature Prep kit 建立台灣地區漢人MPS 之27 個 體染色體STR 資料庫. 32 二、 以ForenSeq™ DNA Signature Prep kit 建立台灣地區漢人MPS 之25 個 Y 染色體STR、7 個X 染色體STR 資料庫. 33 三、 以ForenSeq™ DNA Signature Prep kit 建立台灣地區漢人MPS 之96 個 體染色體SNP 資料庫. 34 第六節 人別鑑定計算. 34 一、 次氯酸鈉裂解DNA 檢體的人別鑑定結果. 35 二、 模擬法醫DNA 檢體的人別鑑定結果. 37 第七節 親緣鑑定計算. 38 一、 次氯酸鈉裂解DNA 檢體的親緣鑑定結果. 39 二、 模擬法醫DNA 檢體的親緣鑑定結.. 40 第五章 討論. 43 第一節 檢體破壞、DNA 萃取及定量. 43 第二節 毛細管電泳法作STR 分析. 44 一、 以毛細管電泳法分析次氯酸鈉裂解DNA 檢體STR. 44 二、 以毛細管電泳法分析模擬法醫DNA 檢體STR. 44 第三節 巨量平行定序(MPS) 法之品質評估. 45 第四節 巨量平行定序(MPS) 法作STR 與SNP 分析. 46 一、 以MPS 分析次氯酸鈉裂解DNA 檢體之STR 及SNP 結果. 46 二、 以MPS 分析模擬法醫DNA 檢體之STR 及SNP 結果. 48 第五節 台灣地區漢人巨量平行定序(MPS) 頻率資料庫. 49 第六節 人別鑑定計算. 50 一、 次氯酸鈉裂解DNA 檢體的人別鑑定. 51 二、 模擬法醫DNA 檢體的人別鑑定. 51 三、 人別鑑定結果比較. 51 第七節 親緣鑑定計算. 52 一、 次氯酸鈉裂解DNA 檢體的親緣鑑定. 52 二、 模擬法醫DNA 檢體的親緣鑑定. 53 三、 親緣鑑定結果比較. 53 第八節 研究限制討論. 54 一、 研究中模擬的DNA 檢體不能完全代表實際情況. 54 二、 每件檢體的分析次數只有一次. 54 三、 需要更好的統計計算方式. 54 第六章 結論與展望. 56 參考文獻. 57 | |
dc.language.iso | zh-TW | |
dc.title | 利用巨量平行定序方法分析裂解DNA之基因型別以進行人別鑑定及親緣鑑定 | zh_TW |
dc.title | Genotyping of degraded DNA using massively parallel sequencing for individual identification and relationship test | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林俊彥(Chun-Yen Lin),謝幸媚(Hsing-Mei Hsieh) | |
dc.subject.keyword | 巨量平行定序,人別鑑定,親緣鑑定,短相連重複序列,單核苷酸多型性,裂解DNA,族群頻率資料庫, | zh_TW |
dc.subject.keyword | massively parallel sequencing,individual identification, relationship test,short tandem repeat,single nucleotide polymorphism,drgraded DNA,population frequency data, | en |
dc.relation.page | 207 | |
dc.identifier.doi | 10.6342/NTU202001464 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2020-07-14 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 法醫學研究所 | zh_TW |
顯示於系所單位: | 法醫學科所 |
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U0001-1307202011503800.pdf 目前未授權公開取用 | 5.35 MB | Adobe PDF |
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