目的 探索过敏性紫癜 （HSP）患儿中是否存在Cosmc基因突变以及该突变与HSP发病的关联性。方法 选取2014年3月至2015年12月确诊为HSP患儿84例为HSP 组，另选取性别和年龄匹配的健康体检者58例为健康对照组。采集两组儿童空腹静脉血5 mL，EDTA 抗凝，根据密度梯度离心法分离得到外周血单个核细胞 （PBMC），提取PBMC中的基因组DNA，应用降落聚合酶链反应 （Touch-down PCR）扩增Cosmc基因全外显子片段，经1% 琼脂糖凝胶电泳鉴定后行测序分析，并进一步分析突变与HSP易感性的关系。结果 测序结果显示HSP 患儿Cosmc 基因存在两个位点的突变，分别为c.393T>A和c.72A>G。c.393T>A和c.72A>G突变位点中各基因型、等位基因在两组间比较差异均无统计学意义，且不受性别影响 （均P>0.05）。结论 HSP 患儿Cosmc基因外显子中存在两个突变位点，且与HSP的发病并无关联。
Abstract
Objective To investigate the presence of Cosmc gene mutation in children with Henoch-Schönlein purpura (HSP) and the association between Cosmc gene mutation and the susceptibility to HSP. Methods Eighty-four children who were diagnosed with HSP between March 2014 and December 2015 were selected as the HSP group. Fiftyeight healthy volunteers matched for age and sex were enrolled as the control group. Fasting venous blood (5 mL) from the two groups was collected in EDTA anticoagulated tubes, followed by the isolation of peripheral blood mononuclear cells (PBMCs) through density gradient centrifugation. Genomic DNA was extracted from PBMCs according to the manufacturer's protocol, and the whole exon region of Cosmc gene was amplified by touch-down polymerase chain reaction (touch-down PCR). The PCR products were identified by 1% agarose gel and sequenced in order to further examine the association between Cosmc gene mutation and the susceptibility to HSP. Results Sequencing results showed two mutations (c.393T>A and c.72A>G) of Cosmc gene in children with HSP. There were no significant differences in the genotype and allele frequencies at the two loci between the HSP and control groups, and this distribution was not associated with sex. Conclusions The mutations c.393T>A and c.72A>G in the exon region of Cosmc gene in children with HSP are not associated with the onset of HSP.
关键词
过敏性紫癜 /
Cosmc 基因 /
基因突变 /
儿童
Key words
Henoch-Schönlein purpura /
Cosmc gene /
Gene mutation /
Child
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Cappellini MD, Fiorelli G. Glucose-6-phosphate dehydrogenase deficiency[J]. Lancet, 2008, 371(9606): 64-74.
[2] Minucci A, Moradkhani K, Hwang MJ, et al. Glucose-6- phosphate dehydrogenase(G6PD) mutations database: review of the "old" and update of the new mutations[J]. Blood Cells MolDis, 2012, 48(3): 154-165.
[3] 陈瑶, 苏跃青, 周进福, 等. Mass-array 芯片技术在葡萄糖-6- 磷酸脱氢酶基因突变位点检测中的应用[J]. 中华检验医学杂志, 2015, 38(12): 822-826.
[4] 严提珍, 钟青燕, 唐宁, 等. 多色探针荧光PCR 熔解曲线法在G6PD 基因突变检测中的临床应用评价[J]. 中华医学遗传学杂志, 2014, 31(2): 156-162.
[5] 林芬, 杨辉, 杨立业. 我国葡萄糖-6- 磷酸脱氢酶缺乏症的分布特征和基因突变[J]. 分子诊断与治疗杂志, 2016, 8(2): 73-77.
[6] Liu WL, Li F, He ZX, et al. Glucose-6-phosphate dehydrogenase qingzhen: Identification of a novel splice mutation (IVS5-1 G>A)[J]. Pediatr Blood Cancer, 2012, 58(5): 825-826.
[7] Jiang W, Yu G, Liu P, et al. Structure and function of glucose- 6-phosphate dehydrogenase-deficient variants in Chinese population[J]. Hum Genet, 2006, 119 (5): 463-478.
[8] Yan JB, Xu HP, Xiong C, et al. Rapid and reliable detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations in Han Chinese using high-resolution melting analysis[J]. J Mol Diagn, 2010, 12(3): 305-311.
[9] Tseng CP, Huang CL, Chong KY, et al. Rapid detection of glucose-6-phosphate dehydrogenase gene mutations by denaturing high-performance liquid chromatography[J]. Clin Biochem, 2005, 38(11): 973-980.
[10] 陈小文, 岳丽杰, 李长钢, 等. 逆转录-PCR- 变性梯度凝胶电泳法检测葡萄糖-6- 磷酸脱氢酶缺乏症[J]. 中华检验医学杂志, 2007, 30(9): 1048-1051.
[11] Myeroff LL, He H, Fink SP, et al. Mutation detection in the TGF-beta receptors and smad genes: RT-PCR and sequencing[J]. Methods Mol Biol, 2000, 142: 139-147.
[12] Liu TC, Yen JS, Shen JS. Rapid molecular diagnosis of hemoglobin variants by RT-PCR of reticulocyte mRNA and direct sequencing[J]. Hemoglobin, 1992, 16(5): 379-388.
[13] 胡淑芬, 罗韬. 从G6PD 缺乏男婴追溯其母亲酶缺陷的漏检情况[J]. 中国优生与遗传杂志, 2013, 21(5): 78-79.
[14] Amini F, Ismail E. 3'-UTR variations and G6PD deficiency[J]. J Hum Genet, 2013, 58(4): 189-194.
基金
上海市浦东新区科技发展基金创新资金（PKJ2012-Y51）。
PDF(1556 KB)
