Detection of Multiple Embryo Growth Factors by Bead-Based Digital Microfluidic Chip

• Main Authors: Tseng, Hsueh-Yang, 曾學洋
• Other Authors: Hsu, Wen-Syang
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/4hrxfb

碩士 === 國立交通大學 === 機械工程系所 === 106 === In 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.