ABO基因突变致A抗原弱表达分析

doi:10.16150/j.1671-2870.2024.06.009

摘要/Abstract

摘要：

目的： 对ABO血清学检测抗原弱表达患者行血型基因分型，分析抗原弱表达的原因。方法： 血清学检测采用微柱凝胶法，血型基因分型采用PCR结合直接测序方法检测患者ABO基因增强子、启动子、第1至第7号外显子以及其相邻的内含子区域，选用软件PolyPhen-2对突变进行核苷酸功能预测和3D结构图绘制。结果： 血清学检测，患者红细胞与抗A抗体反应弱凝集（2+）、与抗B抗体反应凝集（4+），患者血清与A1细胞反应弱凝集（2+），与B细胞和O细胞反应不凝集，其ABO血型血清学表现型为A_weakB，抗球蛋白试验和抗体筛查试验均为阴性。DNA测序提示6号外显子存在c.297 A>G突变，7号外显子存在c.526 C>G、c.657 C>T、c.703 G>A、c.796 C>A、c.803 G>C、c.930 G>A、c.940 A>G突变。查询国际输血协会数据库，c.940 A>G突变导致ABO基因编码糖基转移酶（glycosyl transferase A，GTA）发生p.Lys314Glu突变，从而导致A酶活性减弱，患者基因型为ABO*AW.37/B.01。结论： 本例患者根据ABO血型DNA测序结果发现其7号外显子存在c.940 A>G突变，此突变位点可使编码的氨基酸发生改变即p.Lys314Glu突变，从而导致A抗原弱表达。

关键词: ABO血型, 基因型, 外显子, 抗原弱表达

Abstract:

Objective This study aims to perform blood group genotyping on patients with weak antigen expression detected by ABO serology and analyze the causes of weak antigen expression. Methods Serological detection was carried out by the microcolumn gel method. Blood group genotyping was conducted by PCR combined with direct sequencing. The enhancer, promoter, exons 1-7, and adjacent intron regions of the ABO blood group gene in the patients were tested. The software PolyPhen-2 was used to predict the nucleotide function of mutations and to generate a 3D structural model. Results In one male patient, the red blood cells showed weak agglutination with anti-A antibody (2+) and agglutination with anti-B antibody (4+). The patient’s serum showed weak agglutination with A1 cells (2+), and no agglutination with B cells and O cells. The serological phenotype of the ABO blood group was A_weakB, with both the direct antiglobulin test and antibody screening test being negative. The results of DNA sequencing showed that there were c.297 A>G mutation in exon 6, and c.526 C>G, c.657 C>T, c.703 G>A, c.796 C>A, c.803 G>C, c.930 G>A and c.940 A>G mutations in exon 7. According to the International Blood Transfusion Association database, the c.940 A>G mutation leads to p.Lys314Glu mutation in the ABO gene-encoded glycosyl transferase A (GTA), resulting in decreased A enzyme activity. The patient’s genotype was ABO*AW.37/B.01. Conclusions In this patient, exon 7 of the ABO blood group gene has a c.940 A>G mutation. This mutation site can change the encoded amino acid, namely p.Lys314Glu, resul-ting in weak expression of A antigen.

Key words: ABO blood group, Genotype, Exon, Weak antigen expression

中图分类号:

R457.11

顾玉微, 王成云, 顾萍, 潘秋辉, 王静. ABO基因突变致A抗原弱表达分析[J]. 诊断学理论与实践, 2024, 23(06): 619-623.

GU Yuwei, WANG Chengyun, GU Ping, PAN Qiuhui, WANG Jing. Analysis of weak expression of A antigen induced by ABO gene mutation[J]. Journal of Diagnostics Concepts & Practice, 2024, 23(06): 619-623.

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Primer name	Primer sequence（5′→3′）	Amplification fragment length/bp
Promoter-E1	F:CGGGAGACAGGGTGTCAAG R:TCGGGCGGTAGGTGCTGAA	731
Enhancer	F:CCCCATCCATAGTCAGATAAGC R:AAAGCAGCCAGCCAGTGTT	724
ABO-E2	F:GAGAAGGAGGGTGAGTGATG R:AGTGAGATGAGCCTGTTGC	323
ABO-E3	F:GTCCCAGAACCAAGAGTGA R:TCCAGAGGTATCCAGGTGA	616
ABO-E4	F:AAGACCAACATCCCAAGAA R:GAGCCACAGGAGGAAAGAG	624
ABO-E5	F:TGCTTACCTGCATCCCACGCT R:TCAAGGAAACCGCCCTCTAAT	572
ABO-E6	F:CATTTGCCTCTGGTTGGTTT R:CTGGAGAAGGAGCTGGGTTT	522
ABO-E7	F:TCTGCTGCTCTAAGCCTTCC R:CCCGTTCTGCTAAAACCAAG	878

Serological identification results	Anti-A	Anti-B	A1c	Bc	Oc	DAT	IAT	ABS
患儿	2+	4+	2+	-	-	-	-	-
AB型对照	4+	4+	-	-	-	-	-	-