Study on glucose-6-phosphate dehydrogenase and gene mutation in neonates with glucose-6-phosphate dehydrogenase deficiency

doi:10.3877/cma.j.issn.1673-5250.2020.06.008

Objective

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.

Methods

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.

Results

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).

Conclusions

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.

Key words: Glucosephosphate dehydrogenase deficiency, Neonatal screening, Glucose-6-phosphatedehydrogenase deficiency, Polymerase chain reaction, Quantitative fluorescence PCR method, Fluorescence PCR melting curve analysis, Regional distribution, Infants, newborn

表1 不同性别患儿及其家系成员G6PD活性比较[U/g Hb，M(P₂₅～P₇₅)]

不同性别患儿及其家系成员	例数	G6PD活性	不同性别患儿及其家系成员	例数	G6PD活性	不同性别患儿及其家系成员	例数	G6PD活性
男、女性患儿	121	0.42(0.35～0.67)	男性患儿	103	0.40(0.32～0.53)	女性患儿	18	1.16(0.92～1.46)
患儿父、母亲	139^a	1.17(0.91～1.42)	男性患儿母亲	103	1.12(0.91～1.28)	女性患儿父、母亲	36	1.74(0.69～2.80)
Z值		-9.981	Z值		-10.832	Z值		-2.021
P值		<0.001	P值		<0.001	P值		0.043

表1 不同性别患儿及其家系成员G6PD活性比较[U/g Hb，M(P₂₅～P₇₅)]

不同性别患儿及其家系成员	例数	G6PD活性	不同性别患儿及其家系成员	例数	G6PD活性	不同性别患儿及其家系成员	例数	G6PD活性
男、女性患儿	121	0.42(0.35～0.67)	男性患儿	103	0.40(0.32～0.53)	女性患儿	18	1.16(0.92～1.46)
患儿父、母亲	139^a	1.17(0.91～1.42)	男性患儿母亲	103	1.12(0.91～1.28)	女性患儿父、母亲	36	1.74(0.69～2.80)
Z值		-9.981	Z值		-10.832	Z值		-2.021
P值		<0.001	P值		<0.001	P值		0.043

表2 本组227例G6PD基因突变携带者的230个G6PD基因突位点分布[例数(%)]

G6PD基因突变位点	构成比	G6PD基因突变位点	构成比
1376G>T	71(30.9)	487G>A	5(2.2)
1388G>A	60(26.1)	1004C>A	4(1.7)
95A>G	24(10.4)	1360C>T	4(1.7)
1024C>T	24(10.4)	383T>C	4(1.7)
871G>A	20(8.7)	592C>T	2(0.9)
392G>T	10(4.3)	517T>C	2(0.9)

表2 本组227例G6PD基因突变携带者的230个G6PD基因突位点分布[例数(%)]

G6PD基因突变位点	构成比	G6PD基因突变位点	构成比
1376G>T	71(30.9)	487G>A	5(2.2)
1388G>A	60(26.1)	1004C>A	4(1.7)
95A>G	24(10.4)	1360C>T	4(1.7)
1024C>T	24(10.4)	383T>C	4(1.7)
871G>A	20(8.7)	592C>T	2(0.9)
392G>T	10(4.3)	517T>C	2(0.9)

表3 本组227例患儿及其家系成员G6PD基因突变携带者地区分布[例数(%)]

G6PD基因突变位点	福建省	广西壮族自治区	广东省	江西省	四川省	其他地区
1376G>T	18(41.9)	10(25.6)	6(17.1)	5(14.7)	8(26.7)	13(28.3)
1388G>A	15(34.9)	14(35.9)	5(14.3)	13(38.2)	3(10.0)	8(17.4)
95A>G	1(2.3)	7(17.9)	1(2.9)	1(2.9)	4(13.3)	4(8.7)
1024C>T	1(2.3)	1(2.6)	8(22.9)	3(8.8)	9(30.0)	7(15.2)
871G>A	1(2.3)	2(5.1)	9(25.7)	7(20.6)	1(3.3)	4(8.7)
392G>T	5(11.6)	1(2.6)	1(2.9)	1(2.9)	2(6.7)	3(6.5)
1004C>A	1(2.3)	1(2.6)	1(2.9)	1(2.9)	0(0)	1(2.2)
487G>A	0(0)	1(2.6)	1(2.9)	1(2.9)	1(3.3)	1(2.2)
1360C>T	0(0)	1(2.6)	1(2.9)	1(2.9)	1(3.3)	2(4.3)
383T>C	0(0)	1(2.6)	1(2.9)	0(0)	1(3.3)	2(4.3)
592C>T	1(2.3)	0(0)	1(2.9)	0(0)	0(0)	1(2.2)
517T>C	0(0)	0(0)	0(0)	1(2.9)	0(0)	0(0)
合计	43(100.0)	39(100.0)	35(100.0)	34(100.0)	30(100.0)	46(100.0)

表3 本组227例患儿及其家系成员G6PD基因突变携带者地区分布[例数(%)]

G6PD基因突变位点	福建省	广西壮族自治区	广东省	江西省	四川省	其他地区
1376G>T	18(41.9)	10(25.6)	6(17.1)	5(14.7)	8(26.7)	13(28.3)
1388G>A	15(34.9)	14(35.9)	5(14.3)	13(38.2)	3(10.0)	8(17.4)
95A>G	1(2.3)	7(17.9)	1(2.9)	1(2.9)	4(13.3)	4(8.7)
1024C>T	1(2.3)	1(2.6)	8(22.9)	3(8.8)	9(30.0)	7(15.2)
871G>A	1(2.3)	2(5.1)	9(25.7)	7(20.6)	1(3.3)	4(8.7)
392G>T	5(11.6)	1(2.6)	1(2.9)	1(2.9)	2(6.7)	3(6.5)
1004C>A	1(2.3)	1(2.6)	1(2.9)	1(2.9)	0(0)	1(2.2)
487G>A	0(0)	1(2.6)	1(2.9)	1(2.9)	1(3.3)	1(2.2)
1360C>T	0(0)	1(2.6)	1(2.9)	1(2.9)	1(3.3)	2(4.3)
383T>C	0(0)	1(2.6)	1(2.9)	0(0)	1(3.3)	2(4.3)
592C>T	1(2.3)	0(0)	1(2.9)	0(0)	0(0)	1(2.2)
517T>C	0(0)	0(0)	0(0)	1(2.9)	0(0)	0(0)
合计	43(100.0)	39(100.0)	35(100.0)	34(100.0)	30(100.0)	46(100.0)

表4 实验室和(或)基因诊断对G6PD缺乏症患儿及其家系成员的确诊情况(例)

G6PD缺乏症诊断	患儿			患儿家系成员			合计确诊
G6PD缺乏症诊断	男性	女性	合计	父亲	母亲	合计	合计确诊
实验室诊断	103	18	121	11	111	122	243
基因诊断	95	18	113	11	103	114	227
实验室和(或)基因诊断	103	21	124	11	112	123	247

表4 实验室和(或)基因诊断对G6PD缺乏症患儿及其家系成员的确诊情况(例)

G6PD缺乏症诊断	患儿			患儿家系成员			合计确诊
G6PD缺乏症诊断	男性	女性	合计	父亲	母亲	合计	合计确诊
实验室诊断	103	18	121	11	111	122	243
基因诊断	95	18	113	11	103	114	227
实验室和(或)基因诊断	103	21	124	11	112	123	247

表5 本组179例G6PD基因突变位点前4位携带者的G6PD活性比较[U/g Hb，M(P₂₅～P₇₅)]

G6PD基因突变位点	例数	G6PD活性
1376G>T	71	0.61(0.35～1.15)
1388G>A	60	0.56(0.40～1.13)
95A>G	24	0.73(0.38～1.17)
1024C>T	24	0.99(0.65～1.25)
χ²值		7.642
P值		0.061

表5 本组179例G6PD基因突变位点前4位携带者的G6PD活性比较[U/g Hb，M(P₂₅～P₇₅)]

G6PD基因突变位点	例数	G6PD活性
1376G>T	71	0.61(0.35～1.15)
1388G>A	60	0.56(0.40～1.13)
95A>G	24	0.73(0.38～1.17)
1024C>T	24	0.99(0.65～1.25)
χ²值		7.642
P值		0.061

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