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目的对21-羟化酶缺乏症患儿及其父母的CYP21A2基因进行突变分析,为能有效进行产前诊断提供科学依据。方法收集2013年8月-2015年12月就诊于柳州市妇幼保健院的14例21-羟化酶缺乏症患儿及其父母外周血,提取基因组DNA,采用Sanger测序及多重连接探针扩增技术(MLPA)分析CYP21A2基因的所有外显子及其两端侧翼序列,并通过父母的基因型进行验证,以此为依据进行胎儿的产前诊断。结果 14例先证者中共检出4种突变,不同形式的点突变占89.3%(25/28),分别为c.293-13C>G(46.4%)、c.518T>A(39.2%)、c.737delA(3.6%),缺失突变占10.7%,均为CYP21A2基因外显子1-3缺失。2例产前诊断胎儿均为携带者。结论 CYP21A2基因突变位点c.293-13C>G及c.518T>A是本地区21-羟化酶缺乏症的常见突变。Sanger测序联合多重连接探针扩增技术可以同时检测出CYP21A2基因的点突变和缺失突变,为疾病的确诊与临床表型的判断提供了可靠依据。
Objective To analyze the mutation of CYP21A2 gene in children with 21-hydroxylase deficiency and its parents, and provide a scientific basis for the effective prenatal diagnosis. Methods Peripheral blood samples from 14 patients with 21-hydroxylase deficiency and their parents who were seen at Liuzhou MCH from August 2013 to December 2015 were collected and genomic DNA was extracted and amplified by Sanger sequencing and multiplex ligation (MLPA) analysis of all exons of CYP21A2 gene and its flanking sequences flanked by the genotype of the parents to verify, as a basis for prenatal diagnosis of the fetus. Results Four kinds of mutations were detected in 14 probands, with 89.3% (25/28) in different forms of point mutations, which were c.293-13C> G (46.4%), c.518T> A (39.2%), , C.737delA (3.6%), deletion mutation accounted for 10.7%, all exon 1-3 deletion of CYP21A2 gene. 2 cases of prenatal diagnosis of fetuses are carriers. Conclusion The mutations of CYP21A2 c.293-13C> G and c.518T> A are common mutations of 21-hydroxylase deficiency in this area. Sanger sequencing combined with multiplex ligation probe amplification technology can simultaneously detect point mutations and deletion mutations of CYP21A2 gene, which provides a reliable basis for the diagnosis of the disease and the determination of the clinical phenotype.