2014-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649723摘要：胚胎著床前篩檢及胚胎著床前基因診斷是現今人工生殖科技治療重要的一環，合併胚胎切片、冷凍胚胎及分子診斷技術，可為經常性流產而導致不易受孕，或家族帶有遺傳疾病的孕婦或家庭，於受孕著床前先篩選健康的胚胎再植入母體，提高孕婦受孕的機率及避免下一代帶有家族遺傳疾病。胚胎著床前篩檢目前多利用微陣列比較基因組雜交技術，檢測具有染色體異常或結構異常的胚胎，但此項技術無法區分出平衡轉位的染色體及四倍體，而另外它的限制為解析度平均僅能到達10Mb，對於低於10Mb的微片段缺失則無法偵測，因此篩選出的胚胎仍具有風險。胚胎著床前基因診斷主要針對導致家族遺傳疾病的基因及微衛星進行疾病遺傳點位分析，但此檢驗設計耗時且無法全面性運用於所有個案。並且目前的方法無法同時診斷基因和染色體。次世代定序是目前最新且高通量的定序方法，可完整的檢測及量化全基因體變異，在胚胎診斷方面，目前國際上學者嘗試努力發展，但還未能開始臨床應用，值得我們進一 步的研究。在此三年計晝，我們將研究運用次世代定序於胚胎著床前篩檢及胚胎著床前基 因診斷，計晝第一年目標為利用全基因體定序於胚胎著床前篩檢，分析胚胎細胞23對染 色體是否具有基因劑量改變，染色體轉位等現象。第二年計晝著重於運用外顯子組測序於 胚胎著床前基因診斷，於一次定序實驗即可同時分析家族遺傳點位及遺傳基因之單核甘酸 多型性，分析檢測胚胎是否帶有家族遺傳點位及基因放大不平均或污染等問題。並且發展 可同時檢測基因及染色體的方法。第三年目標將技術實際運用於臨床胚胎著床前篩檢及胚 胎著床前基因診斷，提高孕婦篩檢健康胚胎的效率。<br> Abstract: Preimplantation genetic screening (PGS) and preimplantation genetic diagnosis (PGD)are important treatment modalities in assisted reproductive technology. These techniquescombine embryo biopsy, cryopreservation, and molecular diagnosis and are gradually used inthe prevention of chromosome abnormalities and gene diseases. PGS generally refers to thescreening of chromosomes of embryos for aneuploidy when discussing infertile couples stressedwith issues involving age, repeated in-vitro fertilization failures, and recurrent miscarriages.PGD is generally applied to the diagnosis of a single gene defect of embryos for couples thathave a single gene mutation and want to ensure their offspring free of the disease.Recently, array comparative genome hybridization (aCGH) has been gradually appliedin PGS studies, which examines the entire chromosome complement from the biopsied cells.However, aCGH does not differentiate balanced rearrangements or tetraploidy. And the otherproblem is the resolution, limited in average of 10 megabases (Mb). If the abnormal deletion islower than 10Mb, it would be not detected. PGD is performed for analysis of a familial mutationtogether with linked polymorphic short tandem repeat markers. The development of a customassay for individual patient family is time-consuming and inefficient. In addition, aCGH canonly detect aneuploidy, and conventional mini-sequencing can only detect mutation.Next generation sequencing (NGS) is a developing and promising technique for geneand chromosomal tests that have not been well studied and applied in PGD and PGS. Thisthree-year project focuses on studying NGS in PGD and PGS. In the first year, we will developwhole genome sequencing for PGS for identification of aneuploidy and chromosomal balancedand unbalanced translocation at the finest resolution. In the second year, our approaches areproposed for PGD using whole exome capture technology aiming at detecting familial mutationand single nucleotide polymorphism. We further attempt to perform NGS for both PGS andPGD simultaneously. In the third year, we will apply NGS on PGS and PGD in the clinicalpractice to achieve normal pregnancies.胚胎著床前篩檢胚胎著床前基因診斷次世代定序全基因體定序外顯子組測 序preimplantation genetic screeningpreimplantation genetic diagnosisnext-generation sequencingwhole genome sequencingwhole exome capture