遗传性高铁血红蛋白血症分子诊断方法的研究  

Method for molecular diagnosis of hereditary methemoglobinemia

作　　者：郑德柱[1] 兰风华[1] 谢飞[1] 吴玉水[1] 朱忠勇[1] 

机构地区：[1]南京军区福州总医院全军医学检验中心,福州300025

出　　处：《中华检验医学杂志》2004年第7期415-419,共5页Chinese Journal of Laboratory Medicine

摘　　要：目的 探讨遗传性高铁血红蛋白血症的分子诊断方法。方法 采用RT-PCR和PCR产物直接测序法,对3例遗传性高铁血红蛋白血症患者细胞色素b5还原酶(b5R)cDNA编码区序列进行分析。通过基因组DNA的PCR-限制性酶切或PCR-序列测定,验证cDNA策略所检出的突变。结果 患者A的b5R cDNA在第527位碱基呈T/C杂合状态,第608位碱基呈G/A杂合状态;患者B的b5R cDNA在第170位碱基和第179位碱基均呈G/A杂合状态;患者C的b5R cDNA在第608位碱基呈G/A杂合状态,第791位碱基呈C/T杂合状态。基因组DNA策略与cDNA策略所得结果一致。结论 建立了遗传性高铁血红蛋白血症的分子诊断方法,并在3例患者中发现了3个以复合杂合子形式存在的、新的b5R基因突变。Objective To investigate molecular diagnostic method for hereditary methemoblobinemia. Methods The cDNA coding sequence of NADH-cytochrome b5 reductase (b5R) from 3 patients with hereditary methemoglobinemia was analyzed by direct sequencing of RT-PCR products and the genomic DNA of b5R gene by PCR-restriction endonuclease digestion or PCR-sequencing. Results The b5R cDNA of patient A was T/C heterozygous at nucleotide 527 and G/A heterozygous at nucleotide 608. The b5R cDNA of patient B was G/A heterozygous at both nucleotide 170 and nucleotide 179. The b5R cDNA of patient C was G/A heterozygous at nucleotide 608 and C/T heterozygous at nucleotide 791. Result of genomic DNA analysis was in agreement with that of cDNA approach. Conclusion The method for molecular diagnosis of hereditary methemoglobinemia was established and 3 novel b5R gene mutations were identified in compound heterozygosity in 3 Chinese patients.

关 键 词：遗传性高铁血红蛋白血症 分子诊断 还原酶 细胞色素B5 

分 类 号：R556.7[医药卫生—血液循环系统疾病]