ABO血型基因分型在白血病患儿疑难血型鉴定中的应用

doi:10.3969/j.issn.1673-8640.2015.10.007

摘要/Abstract

摘要： 目的

将基因分型技术应用于白血病患儿的疑难血型鉴定中,有效避免疾病等外在因素的干扰,准确鉴定ABO血型,保障患者的输血安全。

方法

ABO血型血清学鉴定采用微柱凝胶法。提取全血DNA,利用聚合酶链反应(PCR)扩增ABO基因 1~7号外显子、增强子和启动子区序列,进行直接测序,测序结果与血型基因变异位点数据库(Blood Group Antigen Gene Mutation Database)比较,进行ABO血型基因分型。

结果

1例患儿血清学实验结果可能为A亚型,基因分型结果为A102/O01,确认为A型。另1例患儿血清学实验结果可能为B亚型,基因分型结果为B101/O01,确认为B型。2例患者均输注相应血型红细胞制品,无任何不良反应。

结论

该2例患儿因疾病造成ABO血清学鉴定正反定型不符,血型难以鉴定,应用ABO血型基因分型技术则能准确鉴定患者的血型,保证输血安全、有效。

关键词: ABO 血型, 基因型, 表现型, 疑难血型

Abstract: Objective

To identify type discrepancy blood accurately by applying genotyping technique in leukemia children for ABO blood type identification, avoiding the interference of disease and other factors, ensuring transfusion safety.

Methods

Micro-column agglutination was used for ABO blood type serological identification. The DNA was extracted from whole blood of leukemia children. Exon 1-7, enhancer and promoter regions of ABO gene were amplified by polymerase chain reaction(PCR). PCR products were directly sequenced, and the results were compared with those of the Blood Group Antigen Gene Mutation Database to identify ABO genotyping.

Results

The serological result of 1 leukemia children might be subgroup A, genotype A102/O01, so it was identified as A blood type. The another serological result might be subgroup B, genotype B101/O01, so it was identified as B blood type. Red cell products of compatible blood type were transfused to the 2 patients without any adverse reaction.

Conclusions

The results of forward and reverse tests for the 2 children are not accordant because of diseases, which make us hard to identify blood type. However, it helps us with ensuring the transfusion safety and effectivity by ABO genotyping technique.

Key words: ABO blood type, Genotype, Phenotype, Discrepancy blood type

中图分类号:

R446.1

叶星晨, 张帆, 顾玉微, 傅启华, 王静. ABO血型基因分型在白血病患儿疑难血型鉴定中的应用[J]. 检验医学, 2015, 30(10): 993-997.

YE Xingchen, ZHANG Fan, GU Yuwei, FU Qihua, WANG Jing.. Application of ABO genotyping in the identification of type discrepancy blood in children with leukemia[J]. Laboratory Medicine, 2015, 30(10): 993-997.

图/表 4

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引物名称	引物序列(5'-3')	扩增片段长度(bp)
ABO-Promoter-1	F: CGGGAGACAGGGTGTCAAG
	R: TCGGGCGGTAGGTGCTGAA	731
ABO- Promoter-2	F: CCGCAGCTTCACGGGTTCGT
	R: ATGGCTCCGCGTCTGGTCTC	404
ABO-Enhancer	F: CCCCATCCATAGTCAGATAAGC
	R: AAAGCAGCCAGCCAGTGTT	853
ABO-E2	F: GAGAAGGAGGGTGAGTGATG
	R: AGTGAGATGAGCCTGTTGC	323
ABO-E3	F: GTCCCAGAACCAAGAGTGA
	R: TCCAGAGGTATCCAGGTGA	616
ABO-E4	F: AAGACCAACATCCCAAGAA
	R: GAGCCACAGGAGGAAAGAG	636
ABO-E5	F: GAACAACCAGGGACAGAGG
	R: CAAAGGGAAAGAGGACAGC	572
ABO-E6	F: CATTTGCCTCTGGTTGGTTT
	R: CTGGAGAAGGAGCTGGGTTT	521
ABO-E7	F: TCTGCTGCTCTAAGCCTTCC
	R: CCCGTTCTGCTAAAACCAAG	878

引物名称	引物序列(5'-3')	扩增片段长度(bp)
ABO-Promoter-1	F: CGGGAGACAGGGTGTCAAG
	R: TCGGGCGGTAGGTGCTGAA	731
ABO- Promoter-2	F: CCGCAGCTTCACGGGTTCGT
	R: ATGGCTCCGCGTCTGGTCTC	404
ABO-Enhancer	F: CCCCATCCATAGTCAGATAAGC
	R: AAAGCAGCCAGCCAGTGTT	853
ABO-E2	F: GAGAAGGAGGGTGAGTGATG
	R: AGTGAGATGAGCCTGTTGC	323
ABO-E3	F: GTCCCAGAACCAAGAGTGA
	R: TCCAGAGGTATCCAGGTGA	616
ABO-E4	F: AAGACCAACATCCCAAGAA
	R: GAGCCACAGGAGGAAAGAG	636
ABO-E5	F: GAACAACCAGGGACAGAGG
	R: CAAAGGGAAAGAGGACAGC	572
ABO-E6	F: CATTTGCCTCTGGTTGGTTT
	R: CTGGAGAAGGAGCTGGGTTT	521
ABO-E7	F: TCTGCTGCTCTAAGCCTTCC
	R: CCCGTTCTGCTAAAACCAAG	878

样本	单克隆抗A	单克隆抗B	人源性抗A	人源性抗B	单克隆抗H	A1细胞	B细胞	O细胞	直接抗球蛋白实验	间接抗球蛋白实验	抗体筛查
患儿1	±	-	+	-	+	-	+	-	-	-	-
患儿2	-	±	-	+	+	+	-	-	-	-	-
正常A型对照	+	-	+	-	+	-	+	-	-	-	-
正常B型对照	-	+	-	+	+	+	-	-	-	-	-

样本	单克隆抗A	单克隆抗B	人源性抗A	人源性抗B	单克隆抗H	A1细胞	B细胞	O细胞	直接抗球蛋白实验	间接抗球蛋白实验	抗体筛查
患儿1	±	-	+	-	+	-	+	-	-	-	-
患儿2	-	±	-	+	+	+	-	-	-	-	-
正常A型对照	+	-	+	-	+	-	+	-	-	-	-
正常B型对照	-	+	-	+	+	+	-	-	-	-	-

样本	突变位点	基因型
患儿1	261delG、C467T	A102/O01
患儿2	261delG、A297G、C526G、C657T、 G703A、C796A、G830C、G930A	B101/O01