本文選題：支氣管哮喘 + 全外顯子組測序　； 參考：《皖南醫(yī)學院》2017年碩士論文

【摘要】：目的:采用全外顯子組測序技術探尋5q31-33區(qū)中兒童支氣管哮喘的易感基因。方法:從皖南醫(yī)學院第一附屬醫(yī)院等多家醫(yī)療單位收集到支氣管哮喘核心家系18例,從中選取1例母親和兒子均為哮喘患者,而父親正常的家系進行全外顯子組測序;另從120例散發(fā)哮喘兒童中選取20例重度患者進行全外顯子組測序,并針對5q31-33區(qū)進行重點分析,以期找出該區(qū)域內與哮喘相關聯的SNPs位點。結果:1、本研究對一例母親和兒子均為哮喘患者,而父親正常的哮喘核心家系進行全外顯子組測序,通過對測序結果的多步驟篩選分析,最終,在該家系中發(fā)現一個母親和兒子共同擁有的錯義突變位點p.Ala514Thr,并且父親不存在該突變位點,該位點位于PCDH 1基因的外顯子2區(qū)。2、通過對20例重度哮喘患兒的全外顯子組測序結果在5q31-33區(qū)內進行重點分析,我們共發(fā)現了124個在哮喘患者和正常對照者之間存在差異的突變位點。結論:1、PCDH1基因可能是支氣管哮喘的易感基因之一。2、全外顯子組測序是探尋復雜疾病易感基因的有效方法之一,尤其對于罕見突變位點的發(fā)掘能力較高,相比其他方法而言,具有樣本量較少、測序通量更高、測序成本更低等優(yōu)勢。目的:通過對PCDH 1基因的啟動子和五個外顯子區(qū)域進行序列測定,從而尋找PCDH 1基因啟動子和編碼區(qū)的多態(tài)性位點。方法:針對PCDH 1基因的啟動子和五個外顯子設計引物,對所收集的138例哮喘兒童和150例健康對照者進行PCR擴增,并對擴增產物進行sanger測序,將測序結果與NCBI及Hap Map數據庫進行對比,以此確定PCDH 1基因多態(tài)性的分布特征。采用Pearsonχ2檢驗比較各位點基因型和等位基因頻率在哮喘組和對照組的分布。結果:1.本實驗共在PCDH1基因中發(fā)現了五個SNPs位點,其中有4個為新發(fā)現的位點。2.本實驗所檢出的五個SNPs的分布及頻率在哮喘組和對照組之間均未見明顯的統(tǒng)計學差異(P0.05)。結論:本實驗尚未檢出PCDH1基因中與哮喘相關聯的SNPs位點,但由于本實驗樣本量較少和可能存在的人群因素的影響,所以還需進行進一步的驗證。
[Abstract]:Objective: to explore the susceptibility gene of bronchial asthma in children in 5q31-33 region by using total exon sequence analysis. Methods: 18 cases of bronchial asthma nuclear family were collected from the first affiliated Hospital of Southern Anhui Medical College. One case of mother and son were selected as asthma patients, while the father's normal family was sequenced. In addition, 20 patients with severe asthma were selected from 120 children with sporadic asthma to sequence the whole exon group, and to analyze the 5q31-33 region in order to find out the SNPs site associated with asthma in this region. Results: 1. In this study, we sequenced the whole exon group of an asthmatic nuclear family whose mother and son were both asthmatic, and finally analyzed the results by multistep screening. A missense mutation site, p.Ala514Thr, shared by mother and son, was found in the family and did not exist in the father. The locus was located in exon 2 of PCDH 1 gene. The results of total exon group sequencing in 20 children with severe asthma were analyzed in the 5q31-33 region. We found a total of 124 mutation sites with differences between asthmatic patients and normal controls. Conclusion: 1 / 1 PCDH1 gene may be one of the susceptible genes of bronchial asthma. Total exon sequencing is one of the effective methods to explore the susceptible genes of complex diseases, especially for rare mutation sites, compared with other methods. It has the advantages of small sample size, higher sequencing flux and lower sequencing cost. Objective: to search for the polymorphic sites of PCDH 1 gene promoter and coding region by sequencing the promoter and five exon regions of PCDH 1 gene. Methods: primers were designed for the promoter and five exons of PCDH 1 gene. PCR amplification was performed in 138 asthmatic children and 150 healthy controls, and the amplified products were sequenced by sanger. The results of sequencing were compared with NCBI and Hap Map databases to determine the distribution characteristics of PCDH 1 gene polymorphism. Pearson 蠂 2 test was used to compare the distribution of genotype and allele frequencies in asthma group and control group. The result is 1: 1. In this study, five SNPs loci were found in the PCDH1 gene, four of which were newly discovered. There was no significant difference in the distribution and frequency of five SNPs between the asthma group and the control group (P 0.05). Conclusion: the SNPs loci associated with asthma in the PCDH1 gene have not been detected in this study, but due to the small sample size and the influence of possible population factors, further verification is needed.
【學位授予單位】：皖南醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R725.6

【參考文獻】

相關期刊論文 前5條

1 李紅燕;隆紅艷;;兒童咳嗽變異性哮喘氣道炎癥研究[J];長春中醫(yī)藥大學學報;2015年02期

2 柏娟;趙京;申昆玲;向莉;陳愛歡;黃穗;黃英;舒暢;王建生;葉榮偉;;北京、重慶、廣州三城市兒童哮喘患病情況調查[J];中華臨床免疫和變態(tài)反應雜志;2010年04期

3 張和;李昌崇;金龍騰;革麗莎;;支氣管哮喘的遺傳學研究進展[J];中華哮喘雜志(電子版);2010年06期

4 全國兒童哮喘防治協作組;中國城區(qū)兒童哮喘患病率調查[J];中華兒科雜志;2003年02期

5 陳愛歡;李昌崇;趙德育;陳志敏;洪建國;;兒童支氣管哮喘診斷與防治指南[J];中華兒科雜志;2008年10期

相關碩士學位論文 前1條

1 金哲;ORMDL3基因單核苷酸多態(tài)性及表達水平與北京地區(qū)兒童哮喘關聯性研究[D];中國人民解放軍軍事醫(yī)學科學院;2010年

，

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