本文選題：綜合征型先天性心臟病 + 拷貝數(shù)變異　； 參考：《中南大學(xué)》2012年博士論文

【摘要】：目的：探討中國漢族人群中拷貝數(shù)變異與綜合征型先天性心臟病的關(guān)系,發(fā)現(xiàn)其新的致病性拷貝數(shù)變異。同時(shí)盡可能的縮小拷貝數(shù)缺失或者重復(fù)的致病區(qū)間,尋找與先天性心臟病相關(guān)的候選致病基因。 研究對象：2009年4月到2011年8月就診于中南大學(xué)湘雅二醫(yī)院小兒心臟外科的140例綜合征型先天性心臟病患兒。該140例患兒排除了常見的三體綜合征及GATA4,NKX2.5,TBX5和TBXl基因突變。其中,男性患兒75例,女性患兒65例。 研究方法：利用美國QIAGEN公司gDNA抽提試劑盒,按照其標(biāo)準(zhǔn)抽提流程對上述140例患兒進(jìn)行全外周靜脈血gDNA抽提。利用美國illumina公司生產(chǎn)的Human660W-Quad及Human Omni1-quad芯片,采用Array-SNP高通量測序技術(shù)對這其進(jìn)行全基因組CNVs分析。我們從中選出四例國內(nèi)較少報(bào)道的綜合征型患兒gDNA樣本,利用美國Applied Biosystems公司生產(chǎn)的ABI Stepone定量PCR儀實(shí)行SYBR(?) Green I嵌合熒光法實(shí)時(shí)定量PCR,并利用Applied Biosystems的StepOne Software v2.1軟件進(jìn)行數(shù)據(jù)分析驗(yàn)證芯片結(jié)果。 結(jié)果：在140例綜合征型先天性心臟病患兒中共發(fā)現(xiàn)21個(gè)致病性CNVs,其發(fā)生比率約為15%。其中22q11.2微缺失9例,發(fā)生比率約為6.7%,在綜合征型先天性心臟病中最為常見。同時(shí),發(fā)現(xiàn)4例國內(nèi)研究較少的綜合征型先天性心臟病致病性CNVs,包括13q33.1-34,17p11.2缺失各一例及2例11q24-25缺失。對其關(guān)鍵基因進(jìn)行實(shí)施定量PCR,其結(jié)果發(fā)現(xiàn)這4個(gè)致病性CNVs中的關(guān)鍵基因拷貝數(shù)約為正常對照組的0.5倍,與芯片結(jié)果一致。 結(jié)論：1)illumina公司生產(chǎn)的Human660W-Quad及Human Omni1-quad芯片可對綜合征型先天性心臟病患兒致病性CNVs進(jìn)行快速而準(zhǔn)確的分析,并提供可靠的遺傳學(xué)信息； 2)13q33.1-34缺失,17p11.2缺失及11q24-25為綜合征型先天性心臟病的致病性CNVs區(qū)間。 背景：利用美國illumina公司的高通量Array-SNP從140例綜合征型先天性心臟病篩選出21個(gè)致病性CNVs。從這21個(gè)致病性CNVs中選出EFNB2, ZFPM2/FOG2為先天性心臟病的候選致病基因。 目的：本研究旨在探討EFNB2,FOG2兩種候選基因與先天性心臟病的相關(guān)性。 研究對象：2009年4月到2011年8月就診于中南大學(xué)湘雅二醫(yī)院小兒心臟外科的孤立型先天性心臟病患兒。從中選出孤立型房間隔缺損15例,室間隔缺損15例,完全性房室間隔缺損15例,共45例患者進(jìn)行EFNB2基因突變篩選；右室雙出口患兒38例進(jìn)行ZFPM2/FOG2基因突變篩選,上述83例孤立型先天性心臟病gDNA樣本已排除常見的GATA4,NKX2.5,TBX5,TBXl基因突變。 研究方法：利用美國QIAGEN公司gDNA抽提試劑盒,按照其標(biāo)準(zhǔn)抽提流程對上述83例患兒進(jìn)行外周靜脈血gDNA抽提。對EFNB2, ZFPM2/FOG2分別設(shè)計(jì)相應(yīng)的特異性引物擴(kuò)增,利用ABI公司BigDye Terminator v1.1測序試劑盒及其3100測序儀對其PCR擴(kuò)增產(chǎn)物進(jìn)行測序分析。 結(jié)果：在38例孤立型右室雙出口中共發(fā)現(xiàn)5例ZFPM2/FOG2錯(cuò)意突變,其發(fā)生比率約為13.2%。其中c.G1015A(p.V339I)、c.G1831A(p.A611T)、c.A2209G(p.K737E)3例為新發(fā)的錯(cuò)意突變,而另外兩例突變(c.A2107C(p.M703L)及c.C2665G(p.Q889E))常見于先天性膈疝,目前在先天性心臟病中尚無報(bào)道。以上5種突變都未收錄于1000genomes或者the Single Nucleotide Polymor Polymorphism數(shù)據(jù)庫(dbSNP)中。在對45例先天性間隔型缺損進(jìn)行EFNB2突變篩查時(shí)發(fā)現(xiàn)1個(gè)同義突變,余樣本未發(fā)現(xiàn)突變。 結(jié)論：1)ZFPM/FOG2在先天性右室雙出口的發(fā)生中起著重要的作用,然而不能確定其突變與某種類型先天性右室雙出口相關(guān)聯(lián)。 2)EFNB2不是先天性間隔型缺損的候選致病基因,在13q33.1-34區(qū)域中仍存在有其他的候選致病基因。
[Abstract]:Objective : To explore the relationship between copy number variation and syndrome type congenital heart disease in Chinese Han population , and find out its new pathogenic copy number variation .

Object : From April 2009 to August 2011 , 140 children with congenital heart disease were seen from pediatric cardiac surgery in Xiangya Hospital , Central South University . Among these 140 cases , the common three - body syndrome and GATA4 , NKX2.5 , TBX5 and TBXl gene mutations were excluded .

Methods : The whole genome CNVs was extracted from the above 140 cases by using the DNA extraction kit from QIANUS , USA . Using the human 660W - Quad and Human Omni1 - quad chip produced by American Inc . , the whole genome CNVs was analyzed by using the array - SNP high - throughput sequencing technique . Four cases of syndrome - type children with less reported syndrome types were selected from them , SYBR ( ? ) was carried out using ABI Stepone quantitative PCR instrument produced by Applied Biosystems , USA . Real - time quantitative PCR was carried out by Green I chimeric fluorescence method , and the results of the chip were verified by using the software of Applied Biosystems StepOne Software v2.1 .

Results : A total of 21 pathogenic CNVs were found in 140 children with congenital heart disease with a rate of about 15 % . Among them , 22q11.2 was absent in 9 cases , the rate of occurrence was about 6.7 % , the most common in syndrome type congenital heart disease was found .

Conclusion : 1 ) Human660W - Quad and Human Omni1 - quad chip can be used for rapid and accurate analysis of pathogenic CNVs in children with syndrome type congenital heart disease , and provide reliable genetic information .


2 ) The 13q33 . 1 - 34 deletion , 17p11.2 deletion and 11q24 - 25 are the pathogenic CNVs of the syndrome type congenital heart disease .

BACKGROUND : Twenty - one pathogenic CNVs were isolated from 140 syndrome - type congenital heart disease by high - throughput Array - SNP of American company A . EFNB2 and ZFPM2 / FOG2 were selected as candidate pathogenic genes for congenital heart disease .

Objective : To investigate the correlation between EFNB2 and FOG2 candidate genes and congenital heart disease .

Objective : From April 2009 to August 2011 , we visited the isolated congenital heart disease of children in Xiangya Hospital , Central and South University . Fifteen patients with isolated atrial septal defect , 15 ventricular septal defects and 15 complete atrioventricular septal defects were selected , and 45 patients were screened for EFNB2 gene mutation ;
The mutation of the ZFPM2 / FOG2 gene was performed in 38 children with right ventricular double - export , and the above 83 cases of isolated congenital heart disease gDNA were excluded from common GATA4 , NKX2.5 , TBX5 , TBXl gene mutation .

Methods : The gDNA extraction was carried out on the peripheral venous blood of 83 cases of the above - mentioned 83 children by using the gDNA extraction kit of QIAN2 , USA . The corresponding specific primers were designed for EFNB2 and ZFPM2 / FOG2 , and the PCR products were sequenced and analyzed by ABI Company ' s Big Dye Penetration v1.1 sequencing kit and its 3100 sequencer .

Results : Five cases of ZFPM2 / FOG2 missense mutation were found in 38 isolated right ventricular double exits , with a rate of about 13.2 % . Among them , c . G1015A ( p . V339I ) , c . G1831A ( p . A611T ) , c . A2209G ( p . K737E ) were found to be new missense mutations .

Conclusion : 1 ) ZFPM / FOG2 plays an important role in the pathogenesis of congenital right ventricular double exit , however , it cannot be determined that the mutation is associated with some type of congenital right ventricular double - outlet .

2 ) EFNB2 is not a candidate pathogenic gene for congenital septal defect , and other candidate pathogenic genes still exist in the 13q33 . 1 - 34 region .
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：R725.4

【共引文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 玄超;心血管疾病有關(guān)的人血管內(nèi)皮保護(hù)方法及生物標(biāo)志物的研究[D];南開大學(xué);2012年

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本文編號：1973462