探讨葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症新生儿G6PD与其基因突变情况。

2017年8月至2019年4月，在上海市儿童医院新生儿筛查中心进行G6PD缺乏症筛查的新生儿为105 766例，初筛G6PD缺乏症呈阳性患儿为217例，选择其中被召回进行G6PD复筛和(或)基因检测的161例新生儿为研究对象。这161例新生儿均接受G6PD复筛，其中146例接受G6PD基因检测。同时对男性患儿及其母亲、女性患儿及其父母(将男性患儿母亲与女性患儿父母统称为患儿家系成员)进行G6PD活性和基因检测，分析患儿及其家系成员G6PD活性、G6PD基因突变位点地域性分布特点，并分析G6PD活性与不同G6PD基因突变位点的关系。采用Mann-Whitney U检验或Kruskal-Wallis H秩和检验，对不同患儿与其家系成员的G6PD活性，以及不同G6PD基因突变位点患儿及其家系成员的G6PD活性进行比较。本研究研究遵循的程序符合2013年新修订的《世界医学协会赫尔辛基宣言》要求，并与患儿监护人签署临床研究知情同意书。

①本组联合实验室诊断和(或)基因诊断的结果，最终确诊的G6PD缺乏症新生儿为124例(经G6PD活性确诊为121例，G6PD基因检测确诊为113例，二者同时确诊为110例)。②经G6PD活性检测确诊为G6PD缺乏症的121例患儿中，男、女性患儿分别为103、18例。这121例患儿的G6PD活性为0.42 U/g血红蛋白(Hb)(0.35～0.67 U/g Hb)，显著低于其家系成员的1.17 U/g Hb(0.91～1.42 U/g Hb)，男性患儿G6PD活性为0.40 U/g Hb(0.32～0.53 U/g Hb)，显著低于其母亲的1.12 U/g Hb(0.91～1.28 U/g Hb)，女性患儿G6PD活性为1.16 U/g Hb(0.92～ 1.46 U/g Hb)，显著低于其父母的1.74 U/g Hb(0.69～2.80 U/g Hb)，并且上述差异均有统计学意义(Z＝ -9.981、-10.832、-2.021，P<0.001、<0.001、＝0.043)。③本组161例受试儿中，共计146例患儿及其185例家系成员进行G6PD基因检测的结果显示，227例(113例患儿与114例患儿家系成员)携带G6PD基因突变，其中3例为复合型突变，共计检出230个G6PD基因突变位点，前4位为1376G>T、1388G>A、95A>G、1024C>T，占77.8%(179/230)。227例G6PD基因突变携带者的祖籍为福建省、广西壮族自治区、广东省、江西省、四川省及其他地区者分别为43、39、35、34、30与46例，其前2位G6PD基因突变位点分别为1376G>T与1388G>A，1388G>A与1376G>T，871G>A与1024C>T，1388G>A与871G>A，1024C>T与1376G>T，以及1376G>T与1388G>A。④本组前4位G6PD基因突变位点(1376G>T、1388G>A、95A>G、1024C>T)携带者的G6PD活性比较，差异无统计学意义(χ²＝7.642，P＝0.061)。

G6PD活性检测和G6PD基因检测，对G6PD缺乏症患儿都具有诊断意义。G6PD缺乏症患儿及其家系成员G6PD基因突变位点分布具有地域性特点，其G6PD基因突变位点与G6PD活性无明显相关性，临床不能以该病患儿G6PD基因突变位点推测其G6PD活性。

To explore the glucose-6-phosphate dehydrogenase (G6PD) and its gene mutation in G6PD deficiency neonates, and to provide reference for clinical diagnosis of G6PD deficiency infants.

A total of 105 766 newborns screened at Neonatal Screening Center of Shanghai Children′s Hospital from August 2017 to April 2019 were collected. A total of 217 children were initially screened positive for G6PD deficiency, including 161 neonates who were recalled for re-screening for G6PD and/or G6PD gene mutations. And these 161 neonates were selected as research subjects. All of them received re-screening for G6PD, and 146 cases received G6PD gene mutation detection.The male children and their mothers, female children and their parents (the male children′s mothers and female children′s parents were collectively referred as family members of children) received quantitative analysis of G6PD enzyme activity and detection of G6PD gene mutation in children and their family members at the same time. The G6PD enzyme activity, G6PD gene mutation site and its regional distribution characteristics in children and their family members, and the relationship between G6PD enzyme activity and different G6PD gene mutation sites were analyzed. Mann-Whitney U test and Kruskal-Wallis H rank sum test were used for comparisons of G6PD enzyme activity between children and their family members, and relation between G6PD enzyme activity and different sites of G6PD gene mutation. The study was in accordance with the requirements of World Medical Association Helsinki Declaration revised in 2013, and informed consent for clinical research was signed with the children′s guardians.

①Based on results of combined laboratory diagnosis and/or genetic diagnosis, 124 newborns with G6PD deficiency were finally diagnosed (121 cases were diagnosed by G6PD enzyme activity test, 113 cases were diagnosed by G6PD gene mutation detection, and 110 cases were diagnosed by these two methods). ②Among the 121 children diagnosed as G6PD deficiency by G6PD enzyme activity test, 103 were male and 18 were female.G6PD enzyme activity of the 121 children was 0.42 U/g hemoglobin (Hb) (0.35-0.67 U/g Hb), which was significantly lower than 1.17 U/g Hb (0.91-1.42 U/g Hb) of their family members. G6PD enzyme activity of male children was 0.40 U/g Hb (0.32-0.53 U/g Hb), which was significantly lower than 1.12 U/g Hb (0.91-1.28 U/g Hb) of their mothers. G6PD enzyme activity in female children was 1.16 U/g Hb (0.92-1.46 U/g Hb), which was significantly lower than 1.74 U/g Hb (0.69-2.80 U/g Hb) of their parents.All the above differences were statistically significant (Z=-9.981, -10.832, -2.021; P<0.001, <0.001, =0.043). ③Among 161 neonates, 146 cases of children and 185 cases of their family members were tested for G6PD gene mutation, and 227 of them (113 children and 114 family members) carried G6PD gene mutation, among which 3 were complex mutations.A total of 230 G6PD gene mutation sites were detected, and the top four were 1376G>T, 1388G>A, 95A>G, and 1024C>T, accounting for 77.8% (179/230). For 227 cases of G6PD gene mutation carriers, 43, 39, 35, 34, 30 and 46 cases were found to be from Fujian Province, Guangxi Zhuang Autonomous Region, Guangdong Province, Jiangxi Province, Sichuan Province and other regions, respectively, the top two G6PD gene mutation sites in the above regions were 1376G>T and 1388G>A, 1388G>A and 1376G>T, 871G>A and 1024C>T, 1388G>A and 871G>A, 1024C>T and 1376G>T, 1376G>T and 1388G>A, respectively. ④There was no significant difference in G6PD enzyme activity among children and their family members with top four G6PD gene mutation sites 1376G>T, 1388G>A, 95A>G and 1024C>T (χ²=7.642, P=0.061).

G6PD enzyme activity testland G6PD gene mutation detection have diagnostic significance for children with G6PD deficiency. G6PD gene mutation site has the characteristics of regional distribution in children with G6PD deficiency and their family members. There is no significant relationship between G6PD gene mutation site and G6PD enzyme activity, so the G6PD enzyme activity of children with G6PD deficiency could not be predicted by G6PD gene mutation site.