PCR-SSP技术应用于类孟买血型定型的可行性——附2例报告  

The feasibility of identification of para-Bombay blood group with PCR-SSP technology(a report of two cases)

作　　者：许桂芳[1] 梁伟 杨亮 梅传亮 黄丹丹[3] 

机构地区：[1]宁波市第一医院检验科 [2]宁波市中心血站 [3]宁波市第二医院干细胞实验室

出　　处：《中国输血杂志》2014年第5期511-513,共3页Chinese Journal of Blood Transfusion

摘　　要：目的探讨PCR-SSP技术用于类孟买血型快速定型的可行性。方法利用Primer5软件针对A101等位基因cDNA第261、703SNP位点不同的碱基自主设计4对序列特异性引物,对2例血清定型疑似类孟买血型做PCRSSP检测,并对其ABO基因第6、7外显子和FUT1基因做测序分析。结果 2例检测对象基因型分别为B/B、A/O;ABO基因测序分别为B101/B101、A101/O01,FUT1基因测序分别为FUT1基因第547-552位AG纯合缺失(AGAGAG→AGAG),547-552位AG杂合缺失和658位C→T杂合突变。结论 PCR-SSP方法可用于类孟买血型的快速分子初筛,作为1种独立的实验手段弥补血清学定型的不足,提高疑似类孟买血型定型的准确率。Objective To evaluate the feasibility of rapid identification of ABO blood group for para-Bombay phenotype using PCR-SSP technology. Methods The four pairs of special specific primers were designed according to the cDNA nts 261,703 of A101 allele with Primer 5 software. Two cases of suspected para-Bombay phenotype were detected using the PCR-SSP,besides,the Exon6 and Exon7 of ABO gene and FUT1 gene were sequenced by the Sequencing Technology. Results The genotype of ABO blood group obtained from PCR-SSP was B/B, A/O respectively, and the exact genotype showed B101/ B101 ,A101/O01 by sequencing technology respectively;The results indicated that one homozygous mutation was detected by direct sequencing in case 1 ：AG deletion at position 547-552 （AGAGAG→AGAG）. Two heterozygous mutations were detected in case 2：AG deletion at position 547-552 and C to T mutation at position 658. Conclusion The PCR-SSP technology coan discriminate the genotype of ABO blood group gene rapidly for the suspected para-Bombay phenotypes,which work as an independent method to offset the defect of serological method, eventually improve the accuracy of identification of ABO blood group for the suspected para-Bombay phenotypes

关 键 词：序列特异性引物-聚合酶链反应 ABO血型 基因分型 测序 ABO基因 FUT1基因 类孟买血型 

分 类 号：R457.11[医药卫生—治疗学] Q503[医药卫生—临床医学]