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dc.contributor.author曾學洋zh_TW
dc.contributor.author徐文祥zh_TW
dc.contributor.authorTseng, Hsueh-Yangen_US
dc.contributor.authorHsu, Wen-Syangen_US
dc.date.accessioned2018-01-24T07:41:56Z-
dc.date.available2018-01-24T07:41:56Z-
dc.date.issued2017en_US
dc.identifier.urihttp://etd.lib.nctu.edu.tw/cdrfb3/record/nctu/#GT070451004en_US
dc.identifier.urihttp://hdl.handle.net/11536/142231-
dc.description.abstractIVF(In Vitro Fertilization),為目前不孕症之治療方式,其原理為將卵子與精子於體外授精形成胚胎,待其發育成熟後植回母體之技術。臨床植入胚胎時,主要採取多胚胎植入以提升懷孕機率,但是此方法會造成多胞胎與其他副作用。採用選擇性單一胚胎植入能降低多胞胎的發生機率,然而目前在IVF的流程中,胚胎的選擇僅依靠胚胎型態(Morphologic Features)作為選擇植回胚胎的依據。先前的研究指出,胚胎培養液中的胚胎成長因子,能夠作為判斷胚胎發育情形及預測懷孕率之參考,因此藉由量測胚胎成長因子,有機會改善胚胎的選擇效率。目前培養胚胎的方法採用微滴培養法,每顆胚胎僅能取得5-10 μL之胚胎培樣液,然而傳統的免疫分析法樣本需求量高,無法量測單一胚胎培養液中的胚胎成長因子。因此需要開發於低樣本量(5-10 μL),仍能量測到極低濃度(數十pg/mL)之胚胎成長因子的檢測技術。本實驗室所開發之磁珠式數位微流體免疫檢測晶片,樣本需求量僅520 nL,能夠量測單一胚胎培養液中的胚胎成長因子,在先前的研究中,此技術已成功量測到真實鼠胚培樣液中胚胎成長因子Mouse IL-1β。本研究進一步使用此技術,以Human IL-1β及Mouse TNF-α作為檢測目標。對於Human IL-1β建立檢量線,可達偵測極限0.0579 pg/mL,反應時間21分鐘,樣本需求量520 nL之檢測結果;對於Mouse TNF-α建立檢量線,可達偵測極限1.55 pg/mL,反應時間27分鐘,樣本需求量520 nL之檢測結果。本研究亦對於實際人類胚胎培養液中IL-1β濃度檢測,並與植回結果進行驗證,結果顯示培養3-5天胚胎之胚胎培養液中IL-1β濃度變化率增加47.34%以上之胚胎,為成功懷孕之胚胎。此外亦可判定若胚胎發育至第五天,其IL-1β濃度為50 pg/mL以上且濃度變化率增加45%以上時,可能為較良好之胚胎。本研究也顯示,於病患成功懷孕的案例中,其中一顆未被胚胎型態法所選之胚胎,其第5天之IL-1β濃度(60.95 pg/mL)及3-5天濃度變化率(+122.45%),明顯高於另一顆由胚胎型態法所選擇胚胎之IL-1β濃度(51.85 pg/mL)及濃度變化率(+47.34%),因此我們的檢測方法,有機會分辨出僅靠胚胎型態選擇無法分辨出之較佳胚胎。zh_TW
dc.description.abstractIn Vitro Fertilization (IVF) is the current treatment for infertility. In clinical treatment, multiple embryo implantations are adopted to improve the pregnancy rate. However, multiple embryo implantations may cause multiple births and other side effects. The use of selective single embryo implantation reduces the chance of multiple births. At present, the selection of embryos only relies on the morphology features. Embryonic growth factor in the embryonic culture medium can be used as a basis for judging embryonic development and predicting pregnancy rate. Therefore, by measuring the embryonic growth factor have the opportunity to improve the efficiency of embryo selection. However, the current method of culturing the embryo is the microtiter culture method. And the traditional immunoassay sample demand is too high, which is not able to measure the embryonic growth factor in a single embryo culture medium. The sampling requirement of the bead-based digital microfluidic immunoassay developed by our laboratory is only 520 nL, and it is able to measure the embryonic growth factor in a single embryo culture medium. In the previous study, this technique has been successfully measured the embryonic growth factor Mouse IL-1β in the clinical mouse embryo culture medium. This study further used this technique for human IL-1β, the limit of detection is 0.0579 pg/mL, the reaction time is 21 minutes, the sample requirement is 520 nL. For the Mouse TNF-α measurement, the limit of detection is 1.55 pg/mL, the reaction time is 27 minutes, the sample demand is 520 nL. This study also measured the concentration of IL-1β in the human embryo culture medium and verified its data with clinical pregnancy results. The results showed that the embryonic culture medium with the rate of change of IL-1β concentration more than + 47.34% was the embryo of the successful pregnancy. It can also be determined that if the embryo develops to the fifth day when the concentration of IL-1β is more than 50 pg / mL and the change rate of concentration of IL-1β increases by more than 45%, it may be a better embryo. The study also showed that in cases where patients were successfully pregnant, one of the embryos was not selected, its concentration of IL-1β on the 5th day (60.95 pg/mL) and the rate of change of 3-5 days (+ 122.45%) was significantly higher than the selected embryo with concentration of IL-1β (51.85 pg/mL) and the rate of change of 3-5 days (+ 47.34%). Therefore, our detection method has the opportunity to distinguish better embryo which is not able to select only by morphology features.en_US
dc.language.isozh_TWen_US
dc.subject人工體外受精zh_TW
dc.subject胚胎成長因子zh_TW
dc.subject胚胎選擇zh_TW
dc.subjectIn Vitro Fertilizationen_US
dc.subjectEmbryonic growth factoren_US
dc.subjectEmbryo selectionen_US
dc.title磁珠式數位微流體晶片於多重胚胎因子檢測之應用zh_TW
dc.titleDetection of Multiple Embryo Growth Factors by Bead-Based Digital Microfluidic Chipen_US
dc.typeThesisen_US
dc.contributor.department機械工程系所zh_TW
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