Simple PCR detection of haptoglobin gene deletion in anhaptoglobinemic patients with antihaptoglobin antibody that causes anaphylactic transfusion reactions
Simple PCR detection of haptoglobin gene deletion in anhaptoglobinemic patients with antihaptoglobin antibody that causes anaphylactic transfusion reactions
复制标题
DOI：
10.1182/blood.v95.4.1138.004k27_1138_1143
复制
发表时间：
2000-02-15
期刊：
影响因子：
20.3
通讯作者：
Kimura, H
中科院分区：
文献类型：
--
作者：
Koda, Y;Watanabe, Y;Kimura, H
2例无结合珠蛋白血症患者经抗结合珠蛋白抗体检测出现过敏性输血反应，Southern印迹分析证实2例患者为Hp（del）基因缺失型纯合子。我们已经确定了一个病人的基因组DNA使用盒介导的聚合酶链反应（PCR）缺失的连接区。然后，使用PCR从对照个体的基因组DNA扩增从Hp的5'断裂点到启动子区的缺失区。对这些区域的DNA序列分析表明，Hp（del）等位基因的5'断裂点位于Hp外显子1上游5.2kb处，3'断裂点位于结合珠蛋白相关基因外显子5上游52和53 bp之间。5'和3'断裂点侧翼的DNA序列之间没有显著的同源性，除了2-bp（TG）的同一性。为了检测基因频率，我们开发了一种简单的PCR方法来检测基因缺失。我们在157名韩国人、523名日本人和284名中国人中分别发现8、16和17个Hp（del）等位基因，但在101名非洲人和100名欧洲-非洲人中未发现Hp（del）。因此，Hp（del）等位基因纯合子个体的发生率预计在日本人中为1/4000，在韩国人中为1/1500，在中国人中为1/1000。这种发病率高于日本人的伊加缺乏症。结合珠蛋白缺乏和抗结合珠蛋白抗体是亚洲人群输血相关过敏反应的原因，应引起更多关注。
Two anhaptaglobinemic patients showing anaphylactic transfusion reactions by antihaptoglobin antibody were found. Southern blot analysis indicated that 2 patients were homozygous for the deleted allele of the haptoglobin gene (Hp(del)) as reported previously. We have identified the junction region of the deletion from genomic DNA of 1 patient using cassette-mediated polymerase chain reaction (PCR). Then, the deleted region from the 5' breakpoint to the promoter region of the Hp was amplified from genomic DNA of a control individual using PCR. DNA sequence analysis of these regions indicated that the 5' breakpoint of the Hp(del) allele was located 5.2 kilobase (kb) upstream of exon 1 of the Hp and the 3' breakpoint was positioned between 52 and 53 base pair (bp) upstream of exon 5 of the haptoglobin-related gene. There was no significant homology between the DNA sequences flanking the 5' and 3' breakpoints, except for a 2-bp (TG) identity. To examine the gene frequency, we have developed a simple PCR method to detect the gene deletion. We found 8, 16, and 17 Hp(del), alleles in 157 Koreans, 523 Japanese, and in 284 Chinese, respectively, but did not find the Hp(del) in 101 Africans or In 100 European-Africans. The incidence of individuals homozygous for the Hp(del) allele was therefore expected to be 1/4000 in Japanese,1/1500 in Koreans, and 1/1000 in Chinese. This incidence is higher than that of IgA deficiency in Japanese. More attention should be paid on haptoglobin deficiency and antihaptoglobin antibody as the cause of transfusion-related anaphylactic reactions in Asian populations.