产前筛查及多重实时荧光定量PCR快速分子诊断非整倍体技术建立-妇产科学专业论文.docxVIP

广州医学院硕士学位论文产前筛查及多重实时荧光定量 广州医学院硕士学位论文 产前筛查及多重实时荧光定量 PCR 快速分子诊断非整倍体技术的建立 PAGE PAGE 5 Prenatal screening and establishing a rapid diagnosis technique for fetal aneuploidies by multiplex real-time fluorescence relative quantitative Polymerase Chain Reaction Master: Jianzhong Sui Supervisor: Prof. Xiaofang Sun The Third Affiliated Hospital of Guangzhou Medical College;Guangzhou 510150,China. Abstract Objective To explore the foreign software of prenatal screening for Down’s syndrome in China. To establish a multiplex Real-time fluorescence relative quantitative PCR method for diagnosis of Down’s and Edward’s syndrome. Methods By the time-resolved fluoroimmunoassay system, the matemal serum PAPP-A and free ?-hCG of 1517 pregnant women were measured in the first trimester, while the maternal serum AFP and free ?-hCG of 4858 pregnant women were measured in the second trimester,the NT of the first trimester fetus were measured also.Then the first trimester data were evaluated by the software of The First Trimester Screening Program offerred by The Fetal Medicine Foundation (FMF), the second trimester data were evaluated by the software of LifeCycle offerred by PerkinElmer company. Using the technique of multiplex Real-time fluorescence relative quantitative PCR, the sequence of the amyloid precursor protein(APP) gene in the Down’s region of chromosome 21 and of the thymidylate synthetase(TYMS) gene on chromosome 18 were amplified in the same tube. 36 blood samples from normal persons (Group A),15 amniotic fluid samples from normal pregnant women (Group B), 21 samples from Down’s patients (Group C) and 6 samples from Edward’s patients (Group D) were detected.The four group of △CT values were analyzed, we established the △CT value of the normal range: -0.93 ~ -0.03 (Mean 土 3 SD) in this study, then 182 unknown samples were tested by this method. Results 1517 pregnant women were screened in the first-trimester, and 40 cases were regarded as high-risk pregnant women,