罕见Ael亚型的血清学和基因序列分析

Serological and genetic sequence analysis of rare Ael subtypes

目的:探讨A_(el)01亚型血清学和分子机制。方法:采用微柱凝胶法和试管法检测先证者ABO表型,多功能非放射性磷酸盐偶联法检测N-乙酰半乳糖胺基转移酶的反应活性,荧光PCR法进行ABO血型基因分型,Sanger一代测序法对第1~7外显子进行测序。结果:先证者血清学试验检测为A_(el),血清和红细胞上均无N-乙酰半乳糖胺基转移酶活性,ABO血型基因分型结果为AO2,基因测序结果确认为A_(el)01/O02,第1~7外显子测序结果为该样本在A101/O02的基础上发生了239G>A的突变,该突变为首次发现。结论:根据血型血清学结果推测,该突变导致了A抗原弱表达。基因分型预测ABO血型表型,基因测序是直接获得ABO亚型的证据,最终须结合ABO血清学方法来确定其ABO血型。

Objective:To investigate serological and molecular mechanism of A_(el)01 subtype.Methods:ABO phenotype of proband was detected by conventional microcolumn gel method and classical test tube method,reaction activity of N-acetylgalactosaminyltransferase was detected by multifunctional nonradioactive phosphate coupling method,ABO blood group genotyping was performed by fluorescent PCR method,and exons 1~7 were sequenced by Sanger generation sequencing method.Results:Serological test of demonstrant was A_(el),and there was no N-acetylgalactosaminyltransferase activity on serum and red blood cells. ABO blood group genotyping result was AO2,the gene sequencing result was confirmed as A_(el)01/O02,and exon 1~7 sequencing result was 239G>A mutation based on A101/O02. The mutation was discovered for the first time.Conclusion:According to results of blood group serology,this mutation leads to weak expression of A antigen. Genotyping predicts ABO blood group phenotype. Gene sequencing is the direct evidence to obtain ABO subtype. Finally,ABO blood group must be determined in combination with ABO serological methods.