论文部分内容阅读
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 the molecular diagnosis of hereditary methemoglobinemia. Methods The sequence of the coding region of cytochrome b5 reductase (b5R) cDNA in 3 hereditary methemoglobinemia patients was analyzed by RT-PCR and PCR products. Mutations detected by the cDNA strategy were verified by PCR-restriction or PCR-sequencing of genomic DNA. Results The b5R cDNA of patient A showed a T / C heterozygote at position 527 and a G / A heterozygote at position 608. The b5R cDNA of patient B had a mutation at the 170th and 179th bases All showed G / A heterozygous state; in patient C, the base of b5R cDNA was G / A heterozygous at the 608th position, and the 791th base was C / T heterozygous. The genomic DNA strategy is consistent with the cDNA strategy. Conclusion A molecular diagnostic method for hematopoietic hereditary disease was established. Three novel mutations of b5R gene were found in three patients.