荧光定量法与G6PD/6PGD比值法在葡萄糖-6-磷酸脱氢酶缺乏症诊断中的临床价值及基因突变分析  

The clinical value of fluorescence quantitative assay and G6PD/6PGD ratio method in the diagnosis of glucose-6-phosphate dehydrogenase deficiency and gene mutation

作　　者：张潇分 田国力 纪伟 周卓 郭静 王燕敏 Zhang Xiaofen;Tian Guoli;Ji Wei;Zhou Zhuo;Guo Jing;Wang Yanmin(Department of Newborn Screening Center,Shanghai Children′s Hospital,School of Medicine ShanghaiJiao tong University,Shanghai 200040,China)

机构地区：[1]上海市儿童医院,上海交通大学医学院附属儿童医院新生儿筛查中心,上海200040

出　　处：《中华检验医学杂志》2022年第5期522-527,共6页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金(21803009);上海交通大学“交大之星”计划医工交叉研究项目(YG2022QN097);上海市儿童医院院级课题国家自然科学基金培育专项(2021YGZQ09)。

摘　　要：目的探讨荧光定量法与G6PD/6PGD比值法在葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症诊断的临床价值及基因突变类型分析。方法选择2018年6月至2021年3月就诊于上海市儿童医院疑似G6PD缺乏症的1201例(男711例,女490例)患儿,采用荧光定量法、G6PD/6PGD比值法和多色熔解曲线分析法对酶活性、比值和基因突变类型进行检测。分析酶活性、比值与基因突变类型的关系,将荧光定量法和G6PD/6PGD比值法的结果与基因突变结果对比分析,采用受试者工作特征(ROC)曲线评价其诊断效能。结果1201份可疑标本中,检出163份(男135份,女28份),其中荧光定量法检出156例,检出率95.71%,G6PD/6PGD比值法检出140例,检出率85.89%。男性患儿酶活性及比值明显低于女性患儿,差异均具有统计学意义(U=642.5、734.5,P均<0.001)。112例接受基因检测,检出92例,其中半合子突变74例,纯合突变1例,杂合突变15例,复合杂合突变2例。15例杂合突变中,荧光定量法检出11例,检出率73.33%,G6PD/6PGD比值法检出4例,检出率26.67%。基因共检出7个突变位点,所占比例分别为c.1388G>A型32.22%、c.1376G>T型30.00%、c.871G>A型13.33%、c.1024C>T型11.11%、c.95A>G型7.78%、c.487G>A型4.44%、c.392G>T型1.11%。c.1376G>T和c.1024C>T、c.487G>A酶活性差异有统计学意义(P<0.001、0.015);c.1024C>T和c.1388G>A、c.1376G>T、c.871G>A、c.95A>G比值差异均有统计学意义(P=0.017、0.002、0.011、0.013)。ROC曲线分析荧光定量法敏感度100%,特异度95.65%,曲线下面积(AUC)0.972。G6PD/6PGD比值法敏感度100%,特异度94.57%,AUC 0.979。荧光定量法和G6PD/6PGD比值法联合检测时敏感度最高96.7%,特异度最高100%,AUC 0.992。结论与荧光定量法相比,G6PD/6PGD比值法可能无法有效检出女性杂合子;G6PD荧光定量法结合G6PD/6PGD比值法检测有利于减少漏诊,结合基因突变分析,可能提高G6PD患儿诊断率。Objective To investigate the diagnostic value of fluorescence quantitative method and G6PD/6PGD ratio method in glucose-6-phosphate dehydrogenase(G6PD)deficiency and the type of gene mutation.Methods A total of 1201 patients(711 males and 490 females)with suspected G6PD deficiency in Shanghai Children′s Hospital were collected from June 2018 to March 2021.Fluorescence quantification method,G6PD/6PGD ratio method and multicolor melting curve were used to detects enzyme activity,ratio and gene mutation type.Comparison of each index and evaluation of its diagnostic efficiency were performed.Results Among 1201 suspicious samples,163 cases(135 males and 28 females)were finally diagnosed.156 cases were diagnosed by fluorescence quantitative method with a detection rate of 95.71%,and 140 cases were diagnosed by G6PD/6PGD ratio method with a detection rate of 85.89%.enzymatic activity of G6PD and ratio of G6PD/6PGD in male were significantly lower than female,and the differences were statistically significant(U=642.5,734.5,P<0.001).112 cases received G6PD gene mutation detection and 92 cases were diagnosed,74 were hemizygous mutations,1 were homozygous mutations,15 were heterozygous mutations,and 2 were compound heterozygous mutations.Among 15 cases of heterozygous mutations,11 cases were diagnosed by fluorescence quantitative method,the diagnosed rate was 73.33%,4 cases were diagnosed by G6PD/6PGD ratio method,and the diagnosed rate was 26.67%.A total of 7 mutation sites were detected and the proportions were c.1388G>A(32.22%),c.1376G>T(30.00%),c.871G>A(13.33%),c.1024C>T(11.11%).c.95A>G(7.78%),c.487G>A(4.44%),c.392G>T(1.11%).The enzymatic activities of c.1376G>T and c.1024C>T,c.487G>A were statistically significant(P<0.001,0.015);the G6PD/6PGD ratios of c.1024C>T and c.1388G>A,c.1376G>T were statistically significant(P=0.017,0.002,0.011,0.013).Fluorescence quantitative method had sensitivity of 100%,specificity of 95.65%,and the area under the curve(AUC)is 0.972.The sensitivity of the G6PD/6PGD ratio method was 100%,t

关 键 词：葡萄糖-6-磷酸脱氢酶缺乏症 荧光定量法 G6PD/6PGD比值法 多色熔解曲线分析法 基因突变 

分 类 号：R725.8[医药卫生—儿科]