Common Genotypes Screening and Clinical Characteristics of G6PD Deficiency in Dai People of Gestational Age in Yingjiang, Yunnan Province
-
摘要:
目的 分析云南省盈江县傣族人群葡萄糖-6-磷酸脱氢酶（G6PD）缺乏症的筛查结果及基因型分布特征，为其防治提供科学理论依据。 方法 采用荧光斑点法（FST）对盈江县傣族孕龄人群416例样本定量检测G6PD酶活性并用二代测序技术（NGS）进行G6PD基因型检测；对基因型与临床表型进行统计学分析。 结果 使用G6PD酶活性检测416例样本，其阳性率为22.12%，其中男性27.42%，女性17.83%，差异具有统计学意义（P < 0.05）；对416例样本进行NGS检测，检出8种常见基因突变类型：c.1311C > T、c.487G > A、c.1388G > A、c.392G > T、c.1376G > T、c.143T > C、c.1024C > T，c.1311C > T突变在盈江地区携带率为33.17%。NGS和FST检测的阳性率分别为53.13%和22.12%。2种方法的结果一致性较差（Kappa = 0.280）。c.1311C > T突变酶活性多正常，如若不计c.1311C > T突变，2种方法对于男性样本结果一致性较好（Kappa = 0.658）高于女性（Kappa = 0.233）。 结论 云南省盈江县傣族人群G6PD缺乏症携带率高，基因突变主要以c.1311C > T、c.487G > A、c.1388G > A、c.392G > T、c.1376G > T为主，c.1311C > T突变与G6PD酶活性多为正常。采用NGS能同时检测德宏州盈江县傣族常见的突变类型，明显提高检测的阳性率，FST对女性杂合和复合杂合突变均有漏检，对男性半合子突变有较高检出率。 Abstract:Objective To analyze the screening results and genotype distribution characteristics of glucose-6-phosphate dehydrogenase （G6PD） deficiency among Dai people in Yingjiang County, Yunnan Province, and to provide a scientific theoretical basis for its prevention and treatment. Methods The G6PD enzyme activity was quantitatively detected in 416 samples of gestational age population of Dai nationality in Yingjiang County by fluorescent spot test （FST）, and the G6PD genotype was detected by next-generation gene sequencing （NGS） technology. The genotype and clinical phenotype were statistically analyzed. Results The G6PD enzyme activity was detected in 416 samples, and the positive rate was 22.12%, of which 27.42% were male and 17.83% were female, and the difference was statistically significant （P < 0.05）. Eight common gene mutation types were identified: c.1311C > T, c.487G > A, c.1388G > A, c.392G > T, c.1376G > T, c.143T > C, c.1024C > T. The carrier rate of c.1311C > T mutation in Yingjiang area was 33.17%. The positive rates of NGS and FST were 53.13% and 22.12%, respectively, and the results of the two methods were poorly consistent （Kappa = 0.280）. The enzyme activity of the c.1311C > T mutation is mostly normal. If the c.1311C > T mutation is not considered, the results of the two methods are more consistent for male samples （Kappa = 0.658） than for females （Kappa = 0.233）. Conclusions The carrier rate of G6PD deficiency is high in the Dai population in Yingjiang County, Yunnan Province. The gene mutations are mainly c.1311C > T, c.487G > A, c.1388G > A, c.392G > T, and c.1376G > T. c.1311C > T mutation and G6PD enzyme activity are mostly normal. The second-generation sequencing technology can simultaneously detect the common mutation types of the Dai people in Yingjiang County, Dehong Prefecture, and significantly improve the positive rate of detection. The fluorescent spot method may miss both female heterozygous and compound heterozygous mutations, and has a higher detection rate for male hemizygous mutations. -
Key words:
- G6PD deficiency /
- Next-generation sequencing /
- Fluorescent spot test
-
表 1 G6PD酶活性筛查阳性情况统计
Table 1. G6PD enzyme activity screening
组别 筛查数（n） 酶缺乏数（n） 阳性率（%） 男性 186 51 27.42 女性 230 41 17.83 合计 416 92 22.12 
下载: 导出CSV
表 2 德宏州盈江县傣族人群G6PD基因型分布（n）
Table 2. Distribution of G6PD genotype among Dai people in Yingjiang，Dehong （n）
突变类型 男性半合子 女性 总计[n（%）] 杂合子 纯合子 c.1311 C > T 50 51 21 122（55.20） c.487G > A 12 18 4 34（10.82） c.1388G > A 6 12 1 19（8.60） c.392G > T 8 7 1 16（7.24） c.1376G > T 4 2 — 6（2.71） c.143T > C — 2 — 2（0.90） c.1024C > T — 1 — 1（0.45） c.1311 C > T/ c.487G > A — 6 — 6（2.71） c.1311 C > T/ c.1388G > A 2 3 — 5（2.26） c.1311 C > T/ c.1376G > T — 3 — 3（1.36） c.1311 C > T/ c.392G > T 1 — — 1（0.45） c.1311 C > T/ c.487G > A/ c.392G > T — 1 — 1（0.45） c.487G > A/ c.392G > T 1 2 — 3（1.36） c.487G > A/ c.1376G > T — 1 — 1（0.45） c.1388G > A/ c.1376G > T 1 — — 1（0.45） 总计 85 109 27 221（100.00）
下载: 导出CSV
表 3 G6PD基因5种主要突变类型与酶活性筛查情况统计（
$\bar x \pm s $ ）Table 3. Five major mutation types of G6PD gene and enzyme activity screening （
$ \bar x \pm s$ ）突变类型 男性 女性 t P n G6PD酶活性（U/gHb） n G6PD酶活性（U/gHb） c.1311C > T 53 2.81 ± 1.20 85 3.30 ± 1.09 −2.492 0.014* c.487G > A 13 1.08 ± 1.10 32 2.88 ± 1.38 −4.178 0.001* c.1388G > A 9 1.38 ± 0.82 16 2.83 ± 0.88 −4.037 0.001* c.392G > T 10 1.66 ± 1.08 11 2.36 ± 1.08 −1.495 0.151 c.1376G > T 5 0.94 ± 0.78 6 2.30 ± 1.70 −1.638 0.136 *P < 0.05。
下载: 导出CSV
表 4 盈江县傣族孕龄人群2种检测方法G6PD缺乏症检测结果（n）
Table 4. Results of G6PD deficiency detection by two methods in pregnant age of Dai population in Yingjiang （n）
FST检测G6PD缺乏症 NGS检测G6PD缺乏症 合计 阴性 阳性 阴性 182 142 324 阳性 13 79 92 合计 195 221 416
下载: 导出CSV
表 5 盈江县傣族男性孕龄人群2种检测方法G6PD缺乏症检测结果（n）
Table 5. Results of G6PD deficiency detection by two methods in male of pregnant age of Dai population in Yingjiang （n）
FST检测G6PD缺乏症 NGS检测G6PD缺乏症 合计 阴性 阳性 阴性 131 4 135 阳性 19 32 51 合计 150 36 186
下载: 导出CSV
表 6 盈江县傣族女性孕龄人群2种检测方法G6PD缺乏症检测结果（n）
Table 6. Results of G6PD deficiency detection by two methods in female of pregnant age of Dai population in Yingjiang （n）
FST检测G6PD缺乏症 NGS检测G6PD缺乏症 合计 阴性 阳性 阴性 146 43 189 阳性 20 21 41 合计 166 64 230
下载: 导出CSV
表 7 FST检测G6PD酶活性漏诊情况（n）
Table 7. Misdiagnosis of G6PD enzyme activity by FST （n）
突变类型 男性
半合子女性 总计
[n（%）]杂合子 纯合子 c.487G > A — 15 1 16（11.27） c.1311 C > T/ c.487G > A — 3 — 3（2.11） c.487G > A/ c.392G > T 1 1 — 2（1.41） c.1311 C > T/c.487G > A/c.392G > T — 1 — 1（0.70） c.1388G > A 1 12 — 13（9.15） c.1311 C > T/c.1388G > A 1 2 — 3（2.11） c.392G > T — 4 — 4（2.82） c.1376G > T 1 — — 1（0.70） c.1311 C > T/ c.1376G > T — 2 — 2（1.41） c.143T > C — 2 — 2（1.41） c.1311 C > T 36 43 16 95（66.90） 合计 40 85 17 142（100.00）
下载: 导出CSV
-
[1] Lee H Y,Ithnin A,Azma R Z,et al. Glucose-6-phosphate dehydrogenase deficiency and neonatal hyperbilirubinemia:Insights on pathophysiology,diagnosis,and gene variants in disease heterogeneity[J]. Frontiers in Pediatrics,2022,10:875877. doi: 10.3389/fped.2022.875877 [2] Luzzatto L,Ally M,Notaro R. Glucose-6-phosphate dehydrogenase deficiency[J]. Blood,2020,136(11):1225-1240. doi: 10.1182/blood.2019000944 [3] 林芬,杨辉,杨立业. 我国葡萄糖-6-磷酸脱氢酶缺乏症的分布特征和基因突变[J]. 分子诊断与治疗杂志,2016,8(2):73-77. [4] He M,Lin K,Huang Y,et al. Prevalence and molecular study of G6PD deficiency in the Dai and Jingpo ethnic groups in the Dehong Prefecture of the Yunnan province[J]. Human Heredity,2018,83(2):55-64. doi: 10.1159/000489009 [5] 詹小芬,张芹,周露,等. 云南省西双版纳州傣族G6PD缺陷症发生率及突变谱研究[J]. 分子诊断与治疗杂志,2015,7(2):83-86. [6] 章印红,朱宝生,王瑞红,等. 荧光测定法定量测定滤纸片干血斑标本G6PD酶活性的可靠性分析[J]. 中国优生与遗传杂志,2007,15(5):44-45. doi: 10.13404/j.cnki.cjbhh.2007.05.020 [7] Geck R C,Powell N R,Dunham M J. Functional interpretation, cataloging, and analysis of 1,341 glucose-6-phosphate dehydrogenase variants[J]. Am J Hum Genet,2023,110(2):228-239. [8] Djigo O,Ould ASM,Diallo SM,et al. Molecular epidemiology of G6PD genotypes in different ethnic groups residing in Saharan and Sahelian Zones of Mauritania[J]. Pathogens,2021,10(8):931. doi: 10.3390/pathogens10080931 [9] 赵颖,巫静帆,李见群,等. 东莞地区葡萄糖-6-磷酸脱氢酶缺乏症的基因突变筛查[J]. 中华医学遗传学杂志,2018,35(6):840-843. [10] 李頔,卓召振,王怡萌,等. 贵阳地区5486例孕妇G6PD缺乏症基因突变筛查[J]. 贵州医科大学学报,2019,44(7):777-780. [11] 苏震,许力丹,李情情,等. 海南地区人群G6PD基因突变分析[J]. 基因组学与应用生物学,2020,39(7):3208-3212. doi: 10.13417/j.gab.039.003208 [12] Gómez-Manzo S,Marcial-Quino J,Vanoye-Carlo A,et al. Glucose-6-phosphate dehydrogenase:Update and analysis of new mutations around the world[J]. International Journal of Molecular Sciences,2016,17(12):2069. doi: 10.3390/ijms17122069 [13] Liu Z,Yu C,Li Q,Cai R,et al. Chinese newborn screening for the incidence of G6PD deficiency and variant of G6PD gene from 2013 to 2017[J]. Human Mutation,2020,41(1):212-221. doi: 10.1002/humu.23911 [14] Sathupak S,Leecharoenkiat K,Kampuansai J. Prevalence and molecular characterization of glucose-6-phosphate dehydrogenase deficiency in the Lue ethnic group of northern Thailand[J]. Scientific Reports,2021,11(1):2956. doi: 10.1038/s41598-021-82477-w [15] Yang R L,Qian G L,Wu D W,et al. A multicenter prospective study of next-generation sequencing-based newborn screening for monogenic genetic diseases in China[J]. World J Pediatr,2023,19(7):663-673. [16] Chen S,Gao J,Wu Q,et al. Reduction of missed diagnosis of G6PD deficiency in heterozygous females by G6PD/6PGD ratio assay combined with ARMS-PCR[J]. Hum Hered,2022,88(1):1-7. -
