TLR3基因多态性与儿童肠道病毒71型感染的相关性研究

发布时间：2018-01-22 03:37

本文关键词： 肠道病毒71型 Toll样受体3 γ-干扰素基因多态性　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:肠道病毒71型(enterovirus 71,EV71)为手足口病的主要病原体之一,感染后临床表现各异,宿主的遗传背景差异在EV71致病机制中起到重要作用。本项研究探讨TLR3c.1377C/T位点基因多态性与EV71手足口病病情严重程度的关系,分析不同基因表型对EV71感染患病风险的影响。方法:收集2014年9月至2015年10月期间青岛地区的EV71检测阳性的手足口病患儿177例,并记录一般情况(性别、年龄)、临床症状(包括发热持续时间,精神症状、吐泻情况及神经系统、心肺查体的阳性体征)、相关实验室检查(如白细胞计数、C-反应蛋白、肌酸激酶同工酶、谷丙转氨酶和谷草转氨酶),根据《手足口病诊疗指南(2010版)》病情的严重的不同将病例组分为轻症组、重症组;同时收集本地区同时期于两院健康体检的儿童225例作为对照组。提取外周血白细胞基因组DNA,采用改良的多重连接酶反应(i MLDR)技术检测EV71手足口病患儿及正常对照组儿童TLR3c.1377C/T的基因多态性。应用酶联免疫吸附法(ELISA)检测各组血清IFN-γ,IL-4的水平。全部数据均采用SPSS16.0软件进行数据统计处理,以p0.05认为有统计学差异。结果:1.TLR3c.1377C/T位点TT、CT及CC基因型分布频率在EV71感染组(42.37%,44.63%,12.99%)与对照组(51.11%,36.89%,12.00%)比较中未见显著性差异(P0.05),同样T、C等位基因分布频率在EV71感染组(64.69%,35.31%)与对照组(69.56%,30.44%)比较中仍无显著统计学差异(P0.05,OR=1.2,95%CI0.9-1.7)。2.TLR3c.1377C/T位点TT基因型在EV71手足口病重症组(17.95%)的频率高于轻症组(9.09%),其差异有显著统计学差异(P0.05)。TLR3c.1377T等位基因在EV71手足口病重症组(42.95%)的频率明显高于轻症组(29.29%),差异仍有统计学差异(P0.05,OR=1.8,95%CI 1.2-2.8)。3.EV71手足口病患儿的发热持续时间、白细胞计数、C-反应蛋白、血糖等指标在TLR3c.1377C/T各基因型间分布有统计学差异(p0.05),TT基因型患儿以上指标均高于CT基因型及CC基因型携带患儿。而性别、年龄、肌酸激酶同工酶、谷丙转氨酶、谷草转氨酶及病毒载量等指标在各基因型分布频率无显著统计学差异(P0.05)。4.在EV71感染组中,血清IFN-γ水平在TT基因型(102.0±24.2 pg/m L,P0.01)、CT基因型(114.1±26.2 pg/m L,P0.05)手足口病患儿中显著低于CC基因型(135.5±36.8 pg/m L),血清IL-4水平在TT基因型(7.3±1.7 pg/m L,P0.01)、CT基因型(6.4±1.3 pg/m L,P0.05)手足口病患儿中显著高于CC基因型(5.5±1.3 pg/m L)。IFN-γ/IL-4比值在TT基因型(14.2±2.8,P0.001)、CT基因型(18.0±3.1,P0.001)患儿中明显低于CC基因型(24.9±4.7)。5.EV71感染组中,血清IFN-γ、IL-4水平及IFN-γ/IL-4比值与EV71病毒载量间均无明显相关性关系(P0.05)结论:1.TLR3c.1377C/T位点基因多态性可能与EV71感染易感性无关。2.TLR3c.1377TT基因型和CT基因型携带患儿在EV71感染时血清IFN-γ水平明显低于CC基因型,可能更易发展为重症。
[Abstract]:Objective: enterovirus 71 (EV71) is one of the major pathogens of hand, foot and mouth disease (HFMD). The genetic background difference of host plays an important role in the pathogenesis of EV71. This study is to investigate the relationship between TLR3c.1377C/T locus polymorphism and the severity of EV71 HFMD. . To analyze the effect of different gene phenotypes on the risk of EV71 infection. From September 2014 to October 2015, 177 cases of hand-foot-mouth disease with EV71 positive were collected. General conditions (sex, age, clinical symptoms, including duration of fever, mental symptoms, diarrhea and nervous system, positive signs of cardiopulmonary examination, and related laboratory tests, such as white blood cell count) were recorded. C- reactive protein, creatine kinase isoenzyme, alanine aminotransferase and alanine aminotransferase were divided into mild group and severe group according to the severe difference of the condition of hand, foot and mouth disease. At the same time, 225 healthy children in the same period were collected as the control group. The genomic DNA of peripheral blood leukocytes was extracted. Modified multiplex ligase reaction (MLDR). The polymorphism of TLR3c.1377C/T gene in children with hand, foot and mouth disease (EV71) and normal control group was detected by Elisa, and the serum IFN- 纬 was detected by enzyme linked immunosorbent assay (Elisa). The level of IL-4. All the data are processed by SPSS16.0 software. Results: 1. TLR3c.1377C / T locus TTCT and CC genotype frequency were 42.37% in EV71 infection group. There was no significant difference between the control group and the control group (51.1136.89 and 12.00), the same as T. The distribution frequency of C allele in EV71 infection group (64.69) and control group (69.56 and 30.44) still had no significant difference (P0.05). OR=1.2. The frequency of TT genotype at TLR3c.1377C / T locus was higher in EV71 HFMD group than in mild group (P < 0.05). 9.09). There was significant statistical difference in the frequency of P0.05N. TLR3c.1377T allele in severe EV71 hand-foot-mouth disease group (42.95). 29.29. There were still significant differences in the duration of fever and white blood cell count in children with HFMD with CI 1.2-2.8 and 3.EV71. The distribution of C-reactive protein and blood glucose in TLR3c.1377C/T genotypes was significantly different (p0.05). The above indexes in TT genotype were higher than those in CT genotype and CC genotype group, but gender, age, creatine kinase isoenzyme and alanine aminotransferase were higher than those in CT genotype and CC genotype group. There was no significant difference in the distribution frequency of glutamic oxaloacetic transaminase and viral load among different genotypes. The level of serum IFN- 纬 was 102.0 卤24.2 pg/m / L in TT genotype and 114.1 卤26.2 pg/m / L in CT genotype. P0.05) Hand-foot-mouth disease children were significantly lower than CC genotype (135.5 卤36.8 pg/m / L). The serum IL-4 level in TT genotypes was 7.3 卤1.7 pg/m / L and 6.4 卤1.3 pg/m / L in CT genotypes. P0.05) in HFMD children, the ratio of CC genotype was significantly higher than that of CC genotype (5.5 卤1.3 pg/m / L). The ratio of IFN- 纬 / IL-4 in TT genotype was 14.2 卤2.8. P0.001 CT genotype was significantly lower in children with CC genotype (18.0 卤3.1) than in CC genotype (24.9 卤4.7). 5. EV71 infection group. Serum IFN- 纬. There was no significant correlation between IL-4 level, IFN- 纬 / IL-4 ratio and EV71 viral load (P0.05). Conclusion:. 1. Polymorphism of TLR3c.1377C- T locus gene may not be associated with susceptibility to EV71 infection. 2. TLR3c.1377TT genotype and CT genotype carry EV71 susceptibility in children. The serum IFN- 纬 level at the time of infection was significantly lower than that of CC genotype. It is more likely to develop into severe disease.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R725.1

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