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研究生:陳瑞雅
研究生(外文):Jui-Ya Chen
論文名稱:胚胎著床前基因診斷於相互轉位及羅柏森氏轉位之應用
論文名稱(外文):Application of pre-implantation genetic diagnosis for reciprocal and Robertsonian translocations
指導教授:曾麗慧曾麗慧引用關係
指導教授(外文):Li-Hui Tseng
口試委員:李建南陳思原
口試日期:2013-06-27
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:分子醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2013
畢業學年度:101
語文別:中文
論文頁數:75
中文關鍵詞:人工試管嬰兒相互轉位羅伯森氏轉位胚胎著床前基因診斷晶片式比較基因體雜交技術囊胚期胚胎切片植入週期活產率
外文關鍵詞:In Vitro Fertilizationreciprocal translocationRobertsonian translocationpre-implantation genetic diagnosisarray comparative genomic hybridizationblastocyst biopsylive birth rate per cycle
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研究背景:胚胎之染色體非整倍體是人工試管嬰兒失敗、流產最主要原因。在染色體平衡性轉位(相互轉位及羅伯森氏轉位)之夫婦有很高機會產生不平衡的配子,導致試管嬰兒失敗。胚胎著床前基因診斷(pre-implantation genetic diagnosis, PGD)可以檢測染色體的數目有無增減,同時偵測有無大片段的缺失或轉位,篩選正常胚胎,植入母體,增加試管嬰兒之成功機率。以往的研究應用螢光染色原位雜交技術(fluorescent in situ hybridization,FISH)執行PGD,成功率為28.6%。然而FISH診斷率過低,所以本研究希望應用晶片式比較基因體雜交技術(Array comparative genomic hybridization;Array-CGH)及囊胚期胚胎切片提高平衡性轉位夫婦試管嬰兒療程之成功率。
材料及方法:本研究收集17 例攜帶染色體平衡性轉位夫婦共執行23 個cycle的PGD。從2011年1月至2013年3月,個案至台大醫院人工生殖中心,經遺傳諮詢後接受試管嬰兒療程並執行PGD。PGD之胚胎切片採用囊胚期胚胎及Array-CGH,篩選正常胚胎植入。
實驗結果: 本研究有17 例攜帶染色體平衡性轉位夫婦,其中1例是羅伯森氏
轉位、16 例是相互轉位,婦女平均年齡為33.8 歲。
17 例夫妻共執行23 個cycle 的PGD,有四對夫妻共8個週期無胚胎植入,其他13 對夫妻共完成15個週期的治療流程,平均植入1.8 顆胚胎。有9例懷孕,治療週期懷孕率為39.1%(9/23),三位孕婦流產,流產率為13.0%(3/23),植入週期活產率為26.1%(6/23),5 對夫妻共得到6 位健康的寶寶,包含一對雙胞胎,尚有1 對確認懷孕正持續追蹤。臨床結果顯示染色體轉位之夫婦在執行PGD 後的成功治療率為35.3%(6/17 couples)。
結論:應用囊胚期胚胎切片及晶片式比較基因體雜交技術,篩選正常胚胎植入,確實能提高攜帶平衡性轉位夫婦執行人工試管嬰兒之成功率。



Background: The most common reason for In Vitro Fertilization (IVF) failure and abortion is aneuploidy gametes. Translocation carriers (including reciprocal translocation and Robertsonian translocation) have an increased risk of reproductive failure or affected off-springs. Studies have shown that pre-implantation genetic diagnosis (PGD) can detect abnormal chromosome with absence of the large fragment and translocations in embryos. Previous detection method by fluorescent in situ hybridization (FISH ) only had 28.6% of pregnancy rate .We try to verify that the success rate for IVF can be increased by using array comparative genomic hybridization(array-CGH) and blastocyst biopsy.
Material and Methods: The study included 17 patients who underwent 23 cycles of PGD for balance translocation carriers who visited the artificial reproductive center of National Taiwan University Hospital. They performed IVF treatment and PGD from January 2011 to March 2013. PGD used blastocyst biopsy and array-CGH screening.
Results: There are 17 couples with balanced translocation. The average female age is 33.8 years old. One couple had Robertsonian translocation, and 16 couples had reciprocal translocation. 17 patients who underwent 23 cycles of PGD but no balance embryo was found in 8 cycles for four couples. The other13 couples completed 15 cycles of PGD and the average transfer embryo was 1.8 pieces. 9 couples got pregnant and pregnancy rate per cycle was 39.1% (9/23). Three women had miscarriage and abortion rate was 13.0 % (3/23). The live birth rate per cycle was 26.1 % (6/23). Five patients delivered 6 healthy babies including a set of twins and one pregnant couple is confirmed. The study showed that the success rate for couples with PGD was 35.3%(6/17couples)。
Conclusion: Application of blastocyst biopsy and array-CGH can enhance the success rate for balance translocation carriers of In Vitro Fertilization.


目 錄
口試委員會審定書……………………………………………………….ii
誌謝……………………………………………………………………….iii
圖目錄……………………………………………………………………viii
表目錄……………………………………………………………………..ix
中文摘要…………………………………………………………………x
英文摘要………………………………………………………………xii
專有名詞中英文對照表………………………………………………xiv
第一章 緒論……………………………………………………………1
1.1簡介…………………………………………………………………1
1.1.1平衡性轉位………………………………………………………3
1.1.2染色體相互轉位…………………………………………………3
1.1.3染色體羅伯森氏轉位……………………………………………3
1.2 Preimplantation genetic diagnosis;PGD………………………4
1.2.1螢光染色體原位雜交技術(fluorescent in situ hybridization,FISH)…………………………………………………………………………….5
1.2.2晶片式比較基因體雜交技術分析法(Array-CGH)……………….6
1.3台灣人工生殖現況……………………………………………………8
1.4研究動機………………………………………………………………8
第二章 實驗材料與儀器………………………………………………11
2.1實驗材料……………………………………………………………11
2.1.1 24sure 試劑組…………………………………………………11
2.1.2洋菜凝膠電泳試劑………………………………………………11
2.2實驗儀器 …………………………………………………………12
2.2.1 PCR熱循環器 …………………………………………………12
2.2.2水平式電泳槽…………………………………………………12
2.2.3離心機…………………………………………………………12
2.2.4 照相機…………………………………………………………12
2.2.5 Water bath …………………………………………………12
第三章 研究方法………………………………………………………13
3.1參與患者………………………………………………………13
3.2人工生殖………………………………………………………13
3.2.1 IVF/ET:In Vitro Fertilization 體外受精與胚胎移植…13
3.2.2 GIFT:Gamete Intrafallopian Transfer精卵輸卵管植入術…14
3.2.3 ZIFT/TET:Zygote Intrafallopian Transfer/Tubal Embryo Transfer 受精卵/胚胎輸卵管植入術。……………………………14
3.2.4. AID:Artificial Insemination Using Donor’s Semen捐贈精子的人工授精…………………………………………………………14
3.3顯微操作技術……………………………………………………….14
3.3.1.ICSI:Intra-cytoplasmic Sperm Injection卵質內精子注射……14
3.3.2.Assisted Hatching協助孵化 …………………………………..14
3.4胚胎切片(Embryo biopsy)…………………………………………16
3.4.1第一和第二極體切片……………………………………………16
3.4.2胚葉細胞切片……………………………………………………16
3.4.3囊胚期細胞切片…………………………………………………17
3.5晶片式比較基因體雜交技術(Array-CGH)………………………..17
3.5.1 Whole Genome Amplification………………………………18
3.5.2 Array-CGH操作……………………………………………….20
3.6 玻璃化冷凍技術(Vitrification)……………………………21
第四章 實驗結果……………………………………………………..23
4.1 Whole Genome Amplification 結果……………………………..23
4.2 Array-CGH結果…………………………………………………..23
4.3 IVF/ET結果………………………………………………………23
第五章 討論………………………………………………………………28
5.1 關於人工生殖技術…………………………………………………28
5.1.1 IVF/ET:In Vitro Fertilization 體外受精與胚胎移植……………28
5.1.2 ICSI (單一精蟲卵質顯微注射) …………………………………28
5.2關於胚胎切片…………………………………………………………29
5.3關於全基因體放大技術………………………………………………30
5.4關於晶片式比較基因體雜交技術(Array-CGH)……………………31
5.5關於玻璃化冷凍技術…………………………………………………31
5.6關於胚胎著床前基因診斷之風險……………………………………33
5.7關於試管嬰兒著床失敗…………………………………………….33
5.8關於未來展望………………………………………………………35
第六章 醫學倫理與遺傳諮商…………………………………………35
6.1人工生殖法…………………………………………………………35
6.2知情同意……………………………………………………………36
6.3 醫學倫理……………………………………………………………36
6.4遺傳諮詢……………………………………………………………38
第七章 參考文獻………………………………………………………40


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