Lee Tracy Yuen Han. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 168-187). / Abstracts in English and Chinese. / Abstract --- p.ii / 摘要 --- p.v / Acknowledgements --- p.vi / Table of contents --- p.viii / List of tables --- p.xii / List of figures --- p.xiii / List of abbreviations --- p.xiv / Chapter Chapter 1: --- Prenatal diagnosis --- p.1 / Chapter 1.1 --- Historical overview --- p.1 / Chapter 1.1.1 --- Prenatal diagnosis --- p.1 / Chapter 1.1.2 --- Circulating fetal nucleated cells --- p.2 / Chapter 1.1.3 --- Cell-free fetal DNA in maternal plasma --- p.3 / Chapter 1.2 --- Biological characteristics of circulating DNA --- p.4 / Chapter 1.3 --- Origin of circulating DNA --- p.6 / Chapter 1.4 --- Clinical applications of circulating fetal DNA --- p.7 / Chapter 1.5 --- Quantitative aberrations in circulating fetal DNA --- p.9 / Chapter 1.6 --- Epigenetic approach in detecting circulating fetal DNA --- p.11 / Chapter Chapter 2: --- Epigenetics --- p.14 / Chapter 2.1 --- Historical overview --- p.14 / Chapter 2.2 --- Mechanisms of DNA methylation --- p.15 / Chapter 2.3 --- Roles of DNA methylation --- p.17 / Chapter 2.4 --- Aberrations in DNA methylation --- p.20 / Chapter 2.5 --- Epigenetic diagnostic markers --- p.22 / Chapter 2.6 --- Significance of epigenetic markers in noninvasive prenatal diagnosis --- p.23 / Chapter 2.7 --- Aim of thesis --- p.23 / Chapter Chapter 3: --- Materials and methods --- p.25 / Chapter 3.1 --- Preparation of samples --- p.25 / Chapter 3.1.1 --- Collection of placental tissues --- p.25 / Chapter 3.1.2 --- Preparation of plasma and blood cells --- p.25 / Chapter 3.2 --- Nucleic acid extraction --- p.26 / Chapter 3.2.1 --- DNA extraction from placental tissues --- p.26 / Chapter 3.2.2 --- DNA extraction from plasma --- p.26 / Chapter 3.2.3 --- DNA extraction from blood cells --- p.29 / Chapter 3.3 --- Bisulfite genomic sequencing --- p.30 / Chapter 3.3.1 --- Principles of bisulfite modification --- p.30 / Chapter 3.3.2 --- Primer design for bisulfite sequencing --- p.31 / Chapter 3.3.3 --- Bisulfite genomic sequencing --- p.31 / Chapter 3.3.3.1 --- Bisulfite modification --- p.31 / Chapter 3.3.3.2 --- Bisulfite genomic sequencing --- p.32 / Chapter 3.3.4 --- Data and statistical analysis --- p.38 / Chapter 3.4 --- MALDI-TOF Mass Spectrometry (MS) --- p.39 / Chapter 3.4.1 --- Principle of MALDI-TOF MS and MassEXTEND assay --- p.39 / Chapter 3.4.2 --- Methylation-sensitive restriction enzyme digestion and MassEXTEND assay for PDE9A and H19 --- p.41 / Chapter 3.5 --- Quantitative measurements of nucleic acids --- p.44 / Chapter 3.5.1 --- Principles of real-time quantitative PCR --- p.44 / Chapter 3.5.2 --- Methylation-specific qMSP assay for M- and U-PDE9A --- p.47 / Chapter Chapter 4: --- Systematic screening of 8 CGIs in third trimester pregnancy --- p.49 / Chapter 4.1 --- Introduction --- p.49 / Chapter 4.2 --- Methods --- p.50 / Chapter 4.2.1 --- Subjects --- p.50 / Chapter 4.2.2 --- Experimental design --- p.51 / Chapter 4.3 --- Results --- p.54 / Chapter 4.3.1 --- Methylation profile of 8 CGIs in maternal blood cells and paired placental tissues --- p.54 / Chapter 4.3.1.1 --- Methylation profile of PDE9A in third trimester pregnancy --- p.56 / Chapter 4.3.1.2 --- Methylation profile of PPP1R2P2 in third trimester pregnancy --- p.60 / Chapter 4.3.1.3 --- Methylation profile of LOC115376 in third trimester pregnancy --- p.65 / Chapter 4.3.1.4 --- Methylation profile of AM L1 in third trimester pregnancy --- p.71 / Chapter 4.3.1.5 --- Methylation profile of COL6A2 in third trimester pregnancy --- p.78 / Chapter 4.3.1.6 --- Methylation profile of PRDM15 in third trimester pregnancy --- p.82 / Chapter 4.3.1.7 --- Methylation profile of CG1111 in third trimester pregnancy --- p.86 / Chapter 4.3.1.8 --- Methylation profile of CGI121 in third trimester pregnancy --- p.90 / Chapter 4.3.2 --- Methylation profile of regions upstream and downstream of PDE9A in maternal blood cells and paired placental tissues --- p.94 / Chapter 4.3.2.1 --- Methylation profiles of PDE9A_A and PDE9ÁؤB in third trimester pregnancy --- p.96 / Chapter 4.3.2.2 --- Methylation profile of PDE9ÁؤC in third trimester pregnancy --- p.100 / Chapter 4.4 --- Discussion --- p.104 / Chapter Chapter 5: --- "Methylation analysis of PPP1R2P2, PDE9A, PDE9A B and PDE9ÁؤC in first trimester pregnancy" --- p.108 / Chapter 5.1 --- Introduction --- p.108 / Chapter 5.2 --- Methods --- p.109 / Chapter 5.2.1 --- Subjects --- p.109 / Chapter 5.2.2 --- Experimental design --- p.109 / Chapter 5.3 --- Results --- p.110 / Chapter 5.3.1 --- Methylation profile of PPP1R2P2.Region A in first trimester pregnancy. --- p.110 / Chapter 5.3.2 --- Methylation profile of PDE9A in first trimester pregnancy --- p.115 / Chapter 5.3.3 --- Methylation profile of PDE9A B in first trimester pregnancy --- p.119 / Chapter 5.3.4 --- Methylation profile of PDE9A C in first trimester pregnancy --- p.123 / Chapter 5.4 --- Discussion --- p.127 / Chapter Chapter 6: --- "Methylation analysis of PPP1R2P2, PDE9A and PDE9ÁؤB in Trisomy 21 pregnancy" --- p.129 / Chapter 6.1 --- Introduction --- p.129 / Chapter 6.2 --- Methods --- p.130 / Chapter 6.2.1 --- Subjects --- p.130 / Chapter 6.2.2 --- Experimental design --- p.131 / Chapter 6.3 --- Results --- p.131 / Chapter 6.3.1 --- Methylation profile of PPP1R2P2 in trisomy 21 placental tissues --- p.131 / Chapter 6.3.2 --- Methylation profile of PDE9A in trisomy 21 placental tissues --- p.136 / Chapter 6.3.3 --- Methylation profile of PDE9A B in trisomy 21 placental tissues --- p.140 / Chapter 6.4 --- Discussion --- p.144 / Chapter Chapter 7: --- Investigation of imprinting status of PDE9A --- p.145 / Chapter 7.1 --- Introduction --- p.145 / Chapter 7.2 --- Methods --- p.147 / Chapter 7.2.1 --- Subjects --- p.147 / Chapter 7.2.2 --- Experimental design --- p.147 / Chapter 7.3 --- Results --- p.149 / Chapter 7.3.1 --- SNP detection in enzyme digested placental tissues by H19 assay --- p.149 / Chapter 7.3.2 --- SNP detection in enzyme digested placental tissues by PDE9A assay --- p.151 / Chapter 7.4 --- Discussion --- p.153 / Chapter Chapter 8: --- Detection of U-PDE9A DNA sequences in maternal plasma --- p.155 / Chapter 8.1 --- Introduction --- p.155 / Chapter 8.2 --- Methods --- p.156 / Chapter 8.2.1 --- Subjects --- p.156 / Chapter 8.2.2 --- Experimental design --- p.156 / Chapter 8.3 --- Results --- p.157 / Chapter 8.3.1 --- Detection of U-PDE9A DNA sequences in maternal plasma --- p.157 / Chapter 8.3.2 --- Clearance of U-PDE9A DNA sequences from maternal plasma after delivery --- p.157 / Chapter 8.4 --- Discussion --- p.160 / Chapter Chapter 9: --- Conclusion and future perspectives --- p.162 / Chapter 9.1 --- Methylation profiles of CpG islands on chromosome 21q --- p.162 / Chapter 9.2 --- Investigation of imprinting status of PDE9A --- p.164 / Chapter 9.3 --- Development of a universal epigenetic marker --- p.165 / Chapter 9.4 --- Future perspectives --- p.166 / References --- p.168
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325757 |
| Date | January 2006 |
| Contributors | Lee, Tracy Yuen Han., Chinese University of Hong Kong Graduate School. Division of Chemical Pathology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xiv, 187 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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