RT-PCR测序法检测G6PD缺乏症基因突变

发布时间：2018-05-19 23:03

本文选题：GPD缺乏症 + 基因突变　；参考：《中国当代儿科杂志》2016年07期

【摘要】：目的葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症是最常见的遗传性溶血性红细胞酶缺陷病,绝大多数为编码区单碱基突变,现有的多种基因突变检测方法均存在漏检率。该研究拟探讨RT-PCR测序法检测G6PD缺乏症基因突变的可行性。方法将2013年8月至2014年7月的195例贫血查因或体检儿童根据G6PD/6GPD比值(1.00)分为G6PD缺乏组(130例)和对照组(G6PD/6GPD比值≥1.00)65例,优化引物设计及PCR扩增条件,采用RT-PCR测序法对G6PD缺乏组及对照组进行全编码序列分析,基因组DNA测序验证。结果 G6PD缺乏组的基因突变检出率为100%,共检出13种错义突变,包括1种新突变。对照组中28例男性均没有检出错义突变;37例女性中13例检出杂合型错义突变,1例国内外未见报道的纯合型同义突变C1191T,14例C1311T多态位点。对照组的女性标本存在较高的G6PD缺乏症(携带者)漏检率(35%,13/37)。结论 RT-PCR测序法对G6PD基因突变检出率高,具有较大临床诊断价值。
[Abstract]:Objective glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most of which are single base mutations in coding region. The aim of this study was to investigate the feasibility of RT-PCR sequencing for detection of G6PD deficiency gene mutations. Methods 195 children with anemia from August 2013 to July 2014 were divided into G6PD deficiency group (n = 130) and control group (n = 65) according to the G6PD/6GPD ratio of 1.00). The primer design and PCR amplification conditions were optimized. The whole coding sequence of G6PD deficiency group and control group were analyzed by RT-PCR sequencing, and the genomic DNA sequencing was verified. Results the detection rate of gene mutation in G6PD deficiency group was 100. 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 males and 13 out of 37 females were found to have heterozygous missense mutations. One homozygous mutation, C1191T, and 14 C1311T polymorphic loci, which were not reported at home and abroad, were detected. The female samples in the control group had higher G6PD deficiency (carrier) failure rate of 35% or 13 / 37%. Conclusion the detection rate of G6PD gene mutation by RT-PCR sequencing is high and has great clinical diagnostic value.
【作者单位】：重庆医科大学附属深圳市儿童医院血液肿瘤科;重庆医科大学附属深圳市儿童医院儿科研究所;重庆医科大学附属深圳市儿童医院检验科;重庆医科大学附属儿童医院血液肿瘤科;
【基金】：深圳市科技计划项目(医疗卫生类)重点项目(201201004)
【分类号】：R440;R725.5

【相似文献】