荧光PCR熔解曲线法检测广东韶关地区G6PD缺乏症基因突变

Detection of glucose-6-phosphate dehydrogenase(G6PD) deficiency gene mutation by Fluorescence PCR Melting Curve Method in Shaoguan of Guangdong

目的 :采用荧光PCR熔解曲线法检测广东韶关地区G6PD缺乏基因突变,获得基因突变谱并进行方法学应用评价。方法 :采用荧光PCR熔解曲线法试剂盒对613例G6PD缺乏症表型筛查阳性样本,分两管检测中国人群常见的12种G6PD基因突变(95A>G、383T>C、392G>T、487G>A、517T>C、592C>T、871G>A、1004C>A、1024C>T、1360C>T、1376G>T、1388G>A)。结果 :613例标本中检出基因突变536例(男性365例,女171例,占87.44%),其中:1388G>A突变210例(占37.30%),1376G>T突变213例(占37.83%),95A>G突变55例(占9.77%),871G>A突变49例(占8.70%),1024C>T突变18例(占3.20%),1004C>A突变2例(占0.35%),1360C>T突变4例(占0.71%),383T>C突变1例(占0.18%),392G>T突变10例(占1.78%),517T>C突变1例(占0.18%),没有发现487G>A、592C>T突变类型。另外,女性双重杂合或纯合突变分布情况为:1388G>A纯合突变7例,1376G>T纯合突变4例,871G>A纯合突变1例,1388G>A/1376G>T杂合突变4例,1388G>A/871G>A杂合突变2例,1388G>A/95A>G杂合突变1例,1376G>T/1024C>T杂合突变1例,1376G>T/95A>G杂合突变1例,1376G>T/392G>T杂合突变2例。结论 :1388G>A,1376G>T,95A>G,871G>A,1024C>T,1004C>A,1360C>T,383T>C,392G>T,517T>C是韶关地区最常见的基因突变类型,与已报道的中国南方人群突变谱相近;荧光PCR熔解曲线法G6PD缺乏症基因结果准确,可广泛应用于临床基因检测。

ObjectiveGene of G6PD patients with deficiency is tested by Fluorescence PCR Melting Curve Method in Shaoguan of Guangdong to obtain the gene mutation spectrum of G6PD deficiency and evaluate this methodological application. Methods Useing the Fluorescence PCR Melting Curve Method kit to test613 cases of G6PD patients with deficiency who was screened in two part to detect the12 types common G6PD gene mutations in Chinese population (95A>G、383T>C、392G>T、487G>A、517T>C、592C>T、871G>A、1004C>A、1024C>T、1360C>T、1376G>T、1388G>A).Results There were536 cases detected positive in613 cases of specimens (365male,171 female); the percentage was87.44%. Analysising the results of536 cases detected positive:210 cases were1388G>A mutations (37.3%),213 cases were1376G>T mutations (37.83%),55 cases were 95A>G mutations (9.77%),49 cases were871G>A mutations (18.70%), ,18 cases were1024C>T mutations (3.2%),2 cases were1004C>A mutations (0.35%),4 cases were1360C>T mutations (0.71%),1 case was383T>C mutations (0.18%),10 cases were392G>T mutations (1.78%) and1 case was517T>C mutation (0.18%).487G>A、592C>T mutation types were not detected. The distribution of heterozygous or double hybrid mutations for female was that7 cases were1388G>A homozygous mutations,4 cases were1376G>T homozygous mutations,1 case was871G>A homozygous mutations,4 case were1388G>A/1376G>T double hybrid mutations,2 cases were1388G>A /871G>A double hybrid mutations,1 case was1388G>A/95A>G double hybrid mutation,1 case was1376G>T/1024C>T double hybrid mutation,1 case was1376G>T/95A>G double hybrid mutation,2 cases were1376G>T/392G>T double hybrid mutations.Conclusion1388G>A,1376G>T,95A>G,871G>A, 1024C>T,1004C>A,1360C>T,383T>C,392G>T,517T>C are the most common types of gene mutation from Shaoguan G6PD deficiency. . Which is similar to the southern Chinese population mutation spectrum that has been reported. The method based on Fluorescence PCR Melting Curve for detection for the types of genetic mutation of G6PD deficiency is accurate and can be applied to clinical molecular diagnosis widely.