Application of ABO genotyping in the identification of type discrepancy blood in children with leukemia

doi:10.3969/j.issn.1673-8640.2015.10.007

Abstract

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

CLC Number:

R446.1

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.

Figures/Tables 4

References 22

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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