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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 曾麗慧(Li-Hui Teng) | |
dc.contributor.author | Yu-En Chen | en |
dc.contributor.author | 陳羽恩 | zh_TW |
dc.date.accessioned | 2021-06-08T01:45:14Z | - |
dc.date.copyright | 2016-08-26 | |
dc.date.issued | 2016 | |
dc.date.submitted | 2016-08-13 | |
dc.identifier.citation | 參考文獻
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J Forensic Sci. 54(6): 1297-1303. 6. 黎娜, 王華鋒, 曾旭輝 (2015) 唾液酸對精蟲的作用. 中國細胞生物學學報. 37(5):734-739. 7. 葛少欽, 趙崢輝, 張雪倩, 郝媛 (2014) 精蟲表觀遺傳修飾及其在胚胎發育過程中的潛在作用. 遺傳. 36(5):439-446. 8. Joe MG, Gary PS, Carl WS (1990). Zinc-induced Secondary Structure Transitions in Human Sperm protamines. J. Biol. Chem. 265(33): 20667-20672. 9. Diane K. Beebe (1991). Emergency Management of the Adult Female Rape Victim. AAETS. 43(6):2041-6. 10. M Stefanidou, G Alevisopoulos , C Spiliopoulou (2010). Fundamental issues in forensic semen detection Problemas fundamentales en la detección forense de semen. West Indian Medical Journal. 59(3):280-283. 11. Westring CG, Wiuf M, Nielsen SJ, Fogleman JC, Old JB, Lenz C, Reich KA, Morling N (2014). SPERM HY-LITER™ for the identification of spermatozoa from sexual assault evidence. Forensic Sci Int Genet. 12:161-167. 12. H. Yu, E.F. Diamandis (1995). Prostate-specific antigen in milk of lactating women. Chin. Chem. 41:54–58. 13. B.C.M. Pang, B.K.K. Cheung (2007). Identification of human semenogelin in membrane strip test as an alternative method for the detection of semen. Forensic Science International. 169:27–31. 14. Emily S. Boward, Stacey L. Wilson (2013). A comparison of ABAcard p30 and RSID-Semen test kits for forensic semen identification. Journal of Forensic and Legal Medicine. 20:1126-1130. 15. Lamis MN, Amany SM, Manal MI, Dina SAF, Taymor MI (2015). Molecular Biomarkers: The Development of MRNA Multiplex RT-PCR assay for the Definitive Identification of Semen. MOJ Toxicology. Retrieved from http://medcraveonline.com/MOJT/MOJT-01-00025.pdf 16. Erin KH, Jack B (2013). Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis. F1000Res. Retrieved from http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3976110/ 17. Trisha L,Eric Buel (2007). The Use of Real-Time PCR for Forensic Stain Identification. Profiles in DNA.10(1):3–5 18. Catherine MH, Elyse SC, Robin W. Cotton M, Catherine MG (2013). The Effects of Differential Extraction Conditions on the Premature Lysis of Spermatozoa. J Forensic Sci. 58(3):744-752. 19. Schoell WM, Klintschar M, Mirhashemi R, Strunk D, Giuliani A, Bogensberger G, Pertl B (1999). Separation of sperm and vaginal cells based on ploidy, MHC class I, CD45, and cytokeratin expression for enhancement of DNA typing after sexual assault. Cytometry. 36(4):319-323. 20. Mado V, David VH, Filip VN, Dieter D (2008). Automatic detection of spermatozoa for laser capture microdissection. Int J Legal Med. 123(2):169-175. 21. Eisenberg A. (2002). Spermatozoa capture during the differential extraction process for STR typing of sexual assault evidence. National Institute of Justice. Retrieved from https://www.ncjrs.gov/pdffiles1/nij/grants/197532.pdf 22. Xue-Bo Li, Qing-ShanWang, Yu Feng, Shu-Hua Ning, Yuan-Ying Miao, Ye-Quan Wang, Hong-Wei Li. (2014). Magnetic bead-based separation of sperm from buccal epithelial cells using a mon-oclonal antibody against MOSPD3. Int J Legal Med. 10:1007-1014. 23. Anslinger K, Bayer K, Danilov SM, Metzger R (2008). Application of sperm-specific antibodies for the separation of sperm from cell mixtures. Forensic Science International: Genetics Supplement Series. 1(1): 394-395. 24. 劉琳, 陳水琴, 胡志敏, 項林平, 厲坤鵬, 董軍磊, 張爾力, 歐元, 趙興春, 葉健 (2014) 混合斑中精蟲細胞分離方法的應用研究進展. 生命科學研究. 18(1):55-59. 25. Joan MB, Natalie D, Daniel M, Jerome P. Ferrance PL, James PL (2006). Microchip-Based Cell Lysis and DNA Extraction from Sperm Cells for Application to Forensic Analysis. J Forensic Sci. 51(2):266-273. 26. Katie MH, Susan LRB, Jerome PF, Kymberly AF, Katherine AK, James PL (2005). Separation of Sperm and Epithelial Cells in a Microfabricated Device: Potential Application to Forensic Analysis of Sexual Assault Evidence. Anal. Chem. 77:742-749. 27. 歐元, 劉蔚然, 董軍磊, 李彩霞, 靳現霆, 劉琳, 趙興春, 葉健 (2013) 微流控蕊片技術用於精蟲與上皮細胞分離的研究. 中國細胞生物學學報. 35(10):1-6. 28. Voorhees JC, Ferrance JP, Landers JP (2006). Enhanced elution of sperm from cotton swabs via enzymatic digestion for rape kit analysis. J Forensic Sci. 51(3):574-9. 29. Christian Carson, Alex Garvin, Michelle Beckwith, Donna Becherer, Kim Gorman (2011). Purification of Sperm DNA from Vaginal Swabs using the Erase Sperm Isolation Kit. National Institute of Justice. Retrieved from http://erase.ptclabs.com/docs/PosterNIJ2011.pdf | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/19105 | - |
dc.description.abstract | 法醫工作中常遇到需檢測精斑之檢體,然而在實務上以差異萃取法(differential extraction)進行精蟲細胞DNA萃取,卻常面臨到檢體嚴重腐敗、精蟲裂解、大量女性上皮細胞DNA競爭PCR等問題,而導致男性STR DNA 型別鑑驗不佳,精子細胞層呈現混合型別,難以個化嫌犯,故本研究引進核酸酶改良式差異萃取法(Erase sperm isolation kit),收集3組男女標準檢體先進行方法確校,確立方法之重複性、靈敏度、檢測時效,並同時與傳統差異萃取法比較優劣,其結果顯示傳統差異萃取法在精蟲細胞層均呈現混合型別,而核酸酶改良式差異萃取法可精準分離出男性單一型別,模擬48小時後腐敗之檢體,其精蟲細胞層仍可完整分析出男性STR DNA型別,當確認核酸酶改良式差異萃取法之有效性後,本研究收集4組法醫實務案件檢體進行傳統差異萃取法及核酸酶改良式差異萃取法之比較,結果顯示以傳統差異萃取法其精蟲細胞層有3案為男女混合型別、1案仍維持女性單一型別,而以核酸酶改良式差異萃取法4案均可分析出男性單一型別,此外本研究亦同時收集18案實務精斑檢體,其中14案精蟲細胞層同樣可分析出男性單一型別,另外2案則為多人(2名以上男性精蟲細胞)男性混合型別,2案檢體因腐敗等因素仍呈現混合型別,綜上所述,核酸酶改良式差異萃取法相較於傳統差異萃取法,實驗的過程減少細胞洗滌的耗損、避免精蟲細胞提前消化,大幅提高男性精蟲細胞的檢出,並且明確分離出精蟲細胞層男性STR DNA單一型別,以利個化嫌犯,期望將此方法引進國內供法醫及鑑識單位參考,未來面對性侵害、姦殺等案件能夠迅速分型出男性型別,彰顯司法正義,維護社會秩序安寧。 | zh_TW |
dc.description.abstract | It is customary in forensics work to analyze sperm samples. In reality, differential extraction, the extraction of sperm DNA, professionals are often faced with many difficulties. These problems include corrupted samples, sperm autolysis, large amounts of female epithelial DNA competing with that of male sperm DNA in PCR, etc, which produces poor male STR DNA test results wherein the sperm cell layer is mixed, making it hard to narrow down to a more specific suspect.This study collected 3 sets of standard male/female samples to use in the modified-nuclease differential extraction (Erase sperm isolation kit), where they are validated, checked for repeatability and sensitivity, and detected for effectiveness. The results are then compared with that of traditional differential extraction to investigate the advantages and limitations.Results from differential extraction showed mixed sperm cell layer presentation, while Erase sperm isolation kit can accurately extract cell materials of the male gender. In a simulated post 48-hr corrupted sample scenario, the sperm cell layer could still be successfully extracted and completely analyze its male STR DNA type. Once the Erase sperm isolation kit is confirmed for its effectiveness, four sets of forensics case samples were tested using this method as well as the differential extraction method for comparison. Results revealed that with the extraction method, 3 cases were of mixture type and 1 remained female type, while with the erase sperm isolation kit yielded all four cases with male type. This study also collected 18 sperm samples from real-life, forensics cases, 14 of which were of single male type, 2 were of multi-person mixture type, and 2 were corrupted and presented as mixture type. In conclusion, the Erase sperm isolation kit, compared to the traditional differential extraction, eliminated the material loss from cell wash and prevented premature sperm cell lysis, this dramatically increased the likelihood of extracting the male sperm cell material and accurately extracting STR DNA single male type, thus facilitating individualization of the suspect in an ongoing investigation. We hope that this method be utilized in respective domestic forensic units for reference in future cases of sexual assault, rape-kill to rapidly extract the male cell type, bring crimes to justice, and maintain social order and peace in Taiwan. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T01:45:14Z (GMT). No. of bitstreams: 1 ntu-105-P03448005-1.pdf: 1550808 bytes, checksum: 80c3f181065ae428b3328e0f61cf4bc9 (MD5) Previous issue date: 2016 | en |
dc.description.tableofcontents | 目錄
致謝 i 中文摘要 ii 英文摘要 iii 第一章 前言 1 第二章 文獻探討 2 第一節 男性精液成分 第二節 男性精蟲生成 2 第三節 男性精蟲構造 3 第四節 精液篩檢試驗 4 一、 多波域光源 4 二、酸性磷酸酶 4 第五節 精液確認試驗 4 一、 精蟲染色 4 二、 前列腺特異性抗原 5 三、 精囊蛋白 5 四、RNA-based 與MS-base protein assay 5 第六節 精蟲細胞DNA的提取 6 第三章 材料與方法 8 第一節 核酸酶改良式差異萃取法之有效性試驗設計 8 第二節 實務案例研究設計 9 第三節 實驗材料與方法 9 一、 模擬陰道棉棒之標準檢體 9 二、 靈敏度測試樣品製備 9 三、 受試者之標準圖譜檢體 10 四、 法務部法醫研究所實務案件檢體 10 五、 傳統差異萃取法(differential extraction) 10 六、 核酸酶改良式差異萃取法 10 七、 矽管柱萃取法再純化DNA 11 八、 DNA定量 11 九、 聚合酶連鎖反應 11 十、 DNA STR毛細管電泳分析 12 十一、圖譜分析 12 第四章 結果 13 第一節 標準檢體在差異萃取法與核酸酶改良式差異萃取法的比較 13 第二節 核酸酶改良式差異萃取法重複性試驗 13 第三節 核酸酶改良式差異萃取法靈敏度測試 14 第四節 實務案件檢體在差異萃取法與核酸酶改良式差異萃取法的比較 15 第五節 實務案件檢體以核酸酶改良式差異萃取法的評估 16 第五章 討論 17 第一節 傳統差異萃取法與核酸酶差異萃取法比較 17 第二節 核酸酶差異萃取法之靈敏度 18 第三節 特殊案件之評估 18 第四節 研究限制及未來展望 19 第六章 結論 20 第七章 參考文獻 21 第八章 圖表 24 表目錄 表一、精斑篩檢試驗與確認試驗 24 表二、精液成分簡介 24 表三、精蟲生成步驟 24 表四、mRNA-based assay各種檢體檢測標的 25 表五、法務部法醫研究所血清證物組100年~103年精斑檢體統計 26 表六、傳統差異萃取法與核酸酶改良式差異萃取法之比較 26 表七、本研究PCR試劑配置及上機溫度條件 27 表八、標準檢體第1組兩種萃取方法之精蟲細胞層比較 28 表九、標準檢體第2組兩種萃取方法之精蟲細胞層比較 29 表十、標準檢體第3組兩種萃取方法之精蟲細胞層比較 30 表十一、標準檢體第1組核酸酶改良式差異萃取之重複性試驗 31 表十二、標準檢體第2組核酸酶改良式差異萃取之重複性試驗 32 表十三、標準檢體第3組核酸酶改良式差異萃取之重複性試驗 33 表十四、核酸酶改良式差異萃取法針對稀釋精液之靈敏度測試 34 表十五、核酸酶改良式差異萃取法針對檢體時效之靈敏度測試 34 表十六、法醫實務案件兩種萃取方法之定量比較 35 表十七、法醫實務案件兩種萃取方法之圖譜比較 35 表十八、精斑實務檢體18案簡介 36 表十九、精斑實務檢體18案以核酸酶改良式差異萃取法其定量與圖譜分析結果 37 圖目錄 圖一、精蟲生成 38 圖二、精蟲變態逐漸成熟之過程 39 圖三、DTT破壞雙硫鍵示意圖 40 圖四、傳統差異萃取法與核酸酶改良式差異萃取法示意圖 40 圖五、核酸酶改良式差異萃取法實驗步驟 41 圖六、精斑實務檢體18案死亡方式統計 42 圖七、精斑實務檢體18案死者年齡分布 42 圖八、精斑實務檢體18案核酸酶改良式差異萃取法之圖譜分析結果 43 圖九、標準檢體經72小時後上皮細胞與精蟲細胞均裂解呈現混合型圖譜 43 圖十、實務案件裂解後,核酸酶改良式差異萃取法其精蟲細胞層呈混合型別圖譜 44 圖十一、核酸酶改良式差異萃取法萃取兩名以上男性檢體,精蟲細胞層圖譜結果 45 | |
dc.language.iso | zh-TW | |
dc.title | 提昇法醫檢體精斑檢測技術之研究 | zh_TW |
dc.title | The research in improving the forensic identification of semen stain form sexual assault evidences | en |
dc.type | Thesis | |
dc.date.schoolyear | 104-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李俊億(James Chun-I Lee),林俊彥(Chun-Yen Lin) | |
dc.subject.keyword | 精斑鑑定,差異萃取法,核酸?,個化, | zh_TW |
dc.subject.keyword | sperm identification,differential extraction,nuclease,individualization, | en |
dc.relation.page | 45 | |
dc.identifier.doi | 10.6342/NTU201602313 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2016-08-15 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
顯示於系所單位: | 分子醫學研究所 |
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