高通量分子测序技术在男性新生儿G6PD缺乏症筛查中的应用研究  

Application of high-throughput molecular sequencing in the screening of male newborns G6PD deficiency

作　　者：杨金玲[1] 陈大宇[1] 黄丽华[1] 韦江艳 YANG Jinling;CHEN Dayu;HUANG Lihua;WEI Jiangyan(Newborn Disease Screening Center,Affiliated Maternity Hospital of Guangxi University of Science and Technology/Affiliated Children′s Hospital of Guangxi University of Science and Technology/Liuzhou Maternity and Child Health Care Hospital,Liuzhou,Guangxi 545001,China)

机构地区：[1]广西科技大学附属妇产医院/广西科技大学附属儿童医院/广西壮族自治区柳州市妇幼保健院新生儿疾病筛查中心,广西柳州545001

出　　处：《国际检验医学杂志》2021年第23期2881-2884,共4页International Journal of Laboratory Medicine

基　　金：广西壮族自治区柳州市科技重大专项(2018AF10501);广西壮族自治区卫生和计划生育委员会自筹经费科研课题(Z20170532)。

摘　　要：目的探讨高通量分子测序(NGS)技术在男性新生儿葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症筛查中的应用,研究本地区男性新生儿G6PD基因多态性及G6PD酶活性筛查的截断值。方法收集男性新生儿足跟干滤纸血片1092例,采用G6PD酶活性分析和NGS技术进行筛查,采用多色探针荧光聚合酶链反应熔解曲线法进行验证。结果本地区男性新生儿G6PD缺乏症阳性率为11.08%,NGS技术的灵敏度、特异度、阳性预测值和阴性预测值分别为95.04%、100.00%、100.00%和99.39%,与G6PD酶活性分析的灵敏度、特异度、阳性预测值和阴性预测值比较,差异均无统计学意义(P>0.05)。c.1376G>T、c.95A>G、c.1388G>A、c.1024C>T突变为本地区男性新生儿G6PD基因常见的突变类型,其酶活性依次升高;常见突变类型为Ⅱ型和Ⅲ型,以Ⅱ型为主,占85.12%。男性新生儿G6PD酶活性检测截断值<4.24 U/gHb,灵敏度为99.28%,特异度为100.00%,受试者工作特征曲线下面积为0.999。结论NGS技术是男性新生儿G6PD缺乏症筛查的可靠方法,设置合理的G6PD酶活性检测截断值,两种检测方法可以互为补充,可为遗传咨询及临床指导用药提供参考依据。Objective To explore the application of high-throughput molecular sequencing(NGS)technology in the screening of male newborns glucose-6-phosphate dehydrogenase(G6PD)deficiency,to study the G6PD gene polymorphism and the cut-off value of G6PD enzyme activity in male newborns in this area.Methods Collected 1092 cases of male newborns heel dry filter paper blood slices,and used G6PD enzyme activity analysis and NGS technology for screening.The multicolor probe fluorescence polymerase-linked reaction melting curve method was used for verification.Results In this area,the positive rate of G6PD deficiency in male newborns was 11.08%.The sensitivity,specificity,positive predictive value and negative predictive value of NGS technology were 95.04%,100.00%,100.00%and 99.39%,respectively.Compared with the sensitivity,specificity,positive predictive value and negative predictive value of G6PD enzyme activity analysis,the differences were not statistically significant(P>0.05).C.1376G>T,c.95A>G,c.1388G>A,and c.1024C>T mutations were common mutation types in the G6PD gene of male newborns in this area,and the enzyme activity increases sequentially.Common mutation types were typeⅡand typeⅢ.The cut-off value of G6PD enzyme activity detection in male newborns was less than 4.24 U/gHb,the sensitivity was 99.28%,the specificity was 100.00%,and the area under the receiver operating characteristic curve was 0.999.Conclusion NGS technology is a reliable method for screening male newborns for G6PD deficiency,set a reasonable cut-off value for the detection of G6PD enzyme activity,the two detection methods could complement each other,which could provide a reference for genetic counseling and clinical guidance of medication.

关 键 词：葡萄糖-6-磷酸脱氢酶缺乏症 高通量分子测序技术 基因 男性 新生儿 截断值 

分 类 号：R722.1[医药卫生—儿科]