全外显子组测序技术寻找马尔尼菲青霉病的易感基因

发布时间：2018-10-08 07:13

【摘要】：目的：通过外显子组测序技术(whole exome sequencing, WES)从基因水平寻找导致马尔尼菲青霉病(Penicilliosis marneffei, PSM)易感的基因位点,从患者基因水平探讨PSM发病机制。同时,探讨γ干扰素抗体(Anti-interferon-gamma autoantibody)与不合并HIV感染的PSM的相关性。方法：本研究收集2013年至2016年期间于广西医科大学第一附属医院确诊PSM的3例患儿及其父母、60例不合并HIV感染的PSM患者、62健康志愿者,收集其外周静脉血及病史资料。 (1)对3例患儿及其父母的基因组DNA提取、纯化,进行全外显子组测序,通过与人类基因组DNA数据库比对,筛选出SNP (Single nucleotide polymorphism)及Indel (Insertion-deletion)突变位点。通过生物信息学方法在患儿家系内及家系间进行筛选,得到候选基因,与NCBI数据库对比后得到可能导致年幼患儿对PSM的易感基因。(2)采用人γ干扰素抗体检测试剂盒,对不合并HIV感染的PSM患者及健康志愿者检测血清γ干扰素抗体,探讨成人患者对PSM易感的可能原因。结果： (1)外显子组测序平均测序深度为111.66×,质量控制指标均大于90%,测序错误率小于1%,得到72682个SNP位点和6505个Indel位点,通过与已知数据库分析比对、生物信息学分析、手动筛选后得到DOCK8基因突变可能对年幼起病的PSM患儿易感PSM起重要作用。(2)不合并HIV感染的PSM患者与健康志愿者之间γ干扰素检测结果有统计学差异。结论：全外显子组测序技术可用于寻找PSM的易感基因,DOCKS基因可能与幼年患儿对PSM易感相关,在成人患者中,γ干扰素抗体导致的成人起病免疫缺陷综合征(Adult-onset immunodeficiency)可能与PSM易感相关。
[Abstract]:Objective: to explore the gene site of (Penicilliosis marneffei, PSM) susceptibility to penicilliosis marneffei by exon group sequencing technique (whole exome sequencing, WES) at gene level, and to explore the pathogenesis of PSM from the gene level of patients. At the same time, to investigate the correlation between interferon gamma antibody (Anti-interferon-gamma autoantibody) and PSM without HIV infection. Methods: from 2013 to 2016, 3 children with PSM diagnosed in the first affiliated Hospital of Guangxi Medical University and 60 healthy volunteers with PSM without HIV infection were collected and their peripheral venous blood and medical history were collected. The results were as follows: (1) the genomic DNA of 3 children and their parents were extracted, purified and sequenced. The mutation sites of SNP (Single nucleotide polymorphism) and Indel (Insertion-deletion) were screened by comparing with the human genomic DNA database. Candidate genes were obtained by bioinformatics in and between pedigrees, and the susceptibility genes to PSM were obtained by comparing with NCBI database. (2) Human interferon 纬 antibody assay kit was used to detect the susceptibility to PSM in young children. Serum interferon 纬 antibodies were detected in PSM patients without HIV infection and in healthy volunteers to explore the possible causes of susceptibility to PSM in adult patients. Results: (1) the average sequencing depth of exon group was 111.66 脳, the quality control index was more than 90, and the error rate of sequencing was less than 1. 72682 SNP loci and 6 505 Indel loci were obtained. The mutation of DOCK8 gene after manual screening may play an important role in the susceptibility to PSM in young children with PSM. (2) the results of interferon 纬 detection in PSM patients without HIV infection were significantly different from those in healthy volunteers. Conclusion: the whole exon sequence technique can be used to search for the susceptible gene of PSM, docks gene, which may be related to the susceptibility to PSM in young children. In adult patients, the susceptibility to PSM may be associated with the incidence of adult immunodeficiency syndrome (Adult-onset immunodeficiency) caused by interferon 纬 antibody.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R519

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