TLR3基因多态性与新生儿肠道病毒感染

发布时间：2018-03-12 16:05

本文选题：新生儿　切入点：肠道病毒　出处：《浙江大学》2016年硕士论文　论文类型：学位论文

【摘要】：研究背景:每年的8～10月为肠道病毒感染的高发期,婴幼儿是易感者。新生儿肠道病毒感染临床表现轻重不一,且多样化。先天性免疫反应是机体抵抗细菌、病毒和真菌等各种病原微生物入侵的第一道防线,可快速识别病原微生物并对其作出应答。Toll样受体(TLR)是主要的模式识别受体,在先天性免疫应答中识别各种病原微生物的保守结构,与多种细菌和病毒感染性疾病密切相关。TLR3主要负责识别病毒的双链RNA和多聚肌胞苷酸,从而触发炎症反应。近年来的研究发现,TLR3在抗病毒免疫应答中起重要作用,但目前其与肠道病毒感染的关系尚不清楚。本研究通过检测TLR3基因rs3775296,rs3775292,rs3775291,rs3775290,rs13126816,rs5743312共六个位点的基因多态性,探讨TLR3基因多态性与新生儿肠道病毒感染易感性的关系。目的:研究TLR3基因的遗传多态性及其与新生儿肠道病毒感染的关系,探讨肠道病毒的遗传易感性以及细胞因子水平的变化。方法:选择2013年5月一 2014年9月间在浙江大学医学院附属儿童医院新生儿病房住院的47例新生儿肠道病毒感染患儿(病例组)和47例非感染性疾病患儿(对照组)为研究对象。留取其住院期间的血液标本,提取基因组DNA,应用聚合酶链反应(PCR)技术分别检测病例组及对照组TLR3基因rs3775296,rs3775292,rs3775291,rs3775290,rs13126816,rs5743312共六个位点的基因多态性,并利用DNA产物直接测序方法进行序列分析,计算其基因型及等位基因频率;并同时采用流式细胞术检测两组细胞因子(IL-2,IL-4,IL-6,IL-10,INF,IFN-r)水平的变化,并进行统计学分析。结果:TLR3基因rs3775292位点有C和G两种等位基因,其在心电图异常组中的分布频率分别为30%、70%,在心电图正常组中的分布频率分别为8%、92%,心电图异常组与正常组相比,C等位基因和新生儿肠道病毒感染心电图异常的风险有关(P0.05)。进一步分析TLR3基因rs3775292位点的基因型频率在心电图异常组与正常组之间的分布差异无统计学意义(P0.05);TLR3基因rs3775290,rs3775296,rs3775291,rs13126816,rs5743312五个位点的基因型和等位基因频率在心电图异常组与正常组之间的分布差异均无统计学意义(P0.05)。TLR3基因rs3775296,rs3775292,rs3775291,rs3775290,rs13126816,rs5743312 六个位点的基因型和等位基因频率在病例组和对照组之间、脑膜炎组与非脑膜炎组之间、发热3天与≤3天组之间、血小板减少与正常组之间的分布差异均无统计学意义(P0.05)。IL-6、IL-10等细胞因子水平病例组与对照组相比差异有统计学意义(P0.05)。IL-2,IL-4等细胞因子水平病例组与对照组基本持平,TNF,IFN-r等细胞因子水平在病例组虽有增高趋势,但与对照组相比差异均无统计学意义(P0.05)。结论:TLR3 基因 rs3775296,rs3775292,rs3775291,rs3775290,rs13126816,rs5743312六个SNP位点基因多态性可能与新生儿肠道病毒感染发病均无显著关联,亦与新生儿肠道病毒感染脑膜炎发病、发热持续时间、血小板减少病情均无显著关联;rs3775292位点的C等位基因可能与新生儿肠道病毒感染心电图异常的风险有关。新生儿肠道病毒感染时IL-6、IL-10多数有增高,IL-2,IL-4,TNF,IFN-r等细胞因子水平与新生儿肠道病毒感染无显著关联。
[Abstract]:Background: every 8~10 months for the high incidence of intestinal infection, infants and young children are susceptible to neonatal enterovirus infection. Clinical manifestations of varying severity, and diversification. The innate immune response is the first line of defense against bacteria, viruses and fungi and other pathogens, rapid identification of pathogenic microorganisms and make the response of.Toll like receptor (TLR) are pattern recognition receptors, conservative structure recognition of various pathogenic microorganisms in the innate immune response, double stranded RNA is closely related with various bacterial and viral infectious disease.TLR3 responsible for identification of viruses and poly I-C, thereby triggering inflammatory reaction. Research in recent years to found that TLR3 play an important role in antiviral immune response, but its relationship with intestinal infection is not clear. This study through the detection of TLR3 gene rs3775296, rs3775292, RS 3775291, rs3775290, rs13126816, rs5743312 gene polymorphism and six loci were, to investigate the TLR3 gene polymorphism and susceptibility of neonatal enterovirus infection. Objective: the relationship between the infection of genetic polymorphisms of TLR3 gene and neonatal enterovirus, to explore the genetic susceptibility of intestinal virus and changes in cytokine levels. Methods: in May 2013 a September 2014 at the children's Hospital Affiliated to Medical College of Zhejiang University in 47 hospitalized cases of neonatal enterovirus infection (case group) and 47 cases of non infectious diseases (control group) as the research object. The blood samples taken during the stay in hospital, to extract genomic DNA by polymerase chain reaction (PCR) technology were detected in case group and control group TLR3 gene rs3775296, rs3775292, rs3775291, rs3775290, rs13126816, rs5743312 gene polymorphism in six sites, And the sequence was analyzed by direct sequencing method of DNA product, calculate the genotype and allele frequency; and at the same time using flow cytometry in two group of cytokines (IL-2, IL-4, IL-6, IL-10, INF, IFN-r) level, and statistical analysis. Results: TLR3 gene locus rs3775292 and C the two alleles of G, the abnormal ECG group frequency distribution were 30%, 70%, the frequency distribution in normal ECG group were 8%, 92%, abnormal ECG group compared with normal group, the risk of abnormal ECG C allele and neonatal enterovirus infection (P0.05) for further analysis. Differences in genotype frequencies of TLR3 gene rs3775292 locus between abnormal ECG group and normal group had no statistical significance (P0.05); TLR3 gene rs3775290, rs3775296, rs3775291, rs13126816, rs5743312 five locus genotype and allele Because of the difference in frequency distribution between the abnormal ECG group and normal group were not statistically significant (P0.05).TLR3 gene rs3775296, rs3775292, rs3775291, rs3775290, rs13126816, rs5743312 six locus genotype and allele frequencies between case group and control group, between meningitis group and non meningitis group, between the fever for 3 days with less than 3 days, the distribution difference between thrombocytopenia and normal group were not statistically significant (P0.05.IL-6), IL-10 cell factor level in case group compared with the control group, the difference was statistically significant (.IL-2, P0.05) IL-4 cell factor level in the case group and the control group was essentially flat, TNF, IFN-r and other cytokines in case group has increased, but compared with the control group showed no significant difference (P0.05). Conclusion: TLR3 gene rs3775296, rs3775292, rs3775291, rs3775290, rs13126816, rs5743312 six SNP loci There were no significant association of gene polymorphism may be associated with neonatal enterovirus infection and neonatal enterovirus infection meningitis, duration of fever, thrombocytopenia patients showed no significant correlation; rs3775292 loci of the C allele may be associated with neonatal enterovirus infection risk of abnormal ECG. Neonatal enterovirus infection IL-6, IL-10 most there is increased, IL-2, IL-4, TNF, IFN-r etc. but there was no significant association between cytokine levels and intestinal virus infection of the newborn.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R722.1

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