遗传性高铁血红蛋白一家系的基因诊断  

Gene diagnosis of hereditary methemoglobinemia from a Chinese pedigree

作　　者：吴玉水[1] 朱忠勇[1] 王瑶[1] 林运团 陈学香[1] 唐玉钗[1] 

机构地区：[1]福州总医院,福州350025

出　　处：《南京部队医药》1999年第6期10-12,共3页

基　　金：全军八五重点课题基金[(159)C10]

摘　　要：目的 检测Ⅰ型遗传性高铁血红蛋白血症(HM)先证者(L72P,长乐)家系成员的细胞色素b5还原酶(b5R)基因的突变。方法 PCR方法扩增正常人和HM先证者之兄、父、母的b5R基因组DNA片段(含第3和第4外显子);用限制性内切酶Apa Ⅰ分析扩增产物。结果PCR扩增产物用Apa Ⅰ酶切后,正常人为649和471 bp;先证者之兄为649、440和31bp,表明存在b5R基因第72密码子纯合子突变:其父、母均为649、471、440和31bp,表明存在b5R基因第72密码子杂合子突变。结论PCR结合Apa Ⅰ内切酶分析可用于b5R基因第72密码子突变的检测;PCR-RFLP方法是检测人群中b5R基因杂合子突变的简捷有效的方法。To detect mutations in cytochrome b5 reductase (b5R) gene from familial members of a propositus (L72P,Changle) with recessive congenital methemoglobinemia type I . Methods Genomic DNA fragments (1120bp) including exon 3 and 4 of b5R gene from normal controls, propositus' brother and parents were amplified by polymerase chain reaction (PCR); PCR products were then restricted by Apa I endonuclease. Results Fragments of 649 and 471bp were generated in the subjects of normal control. Fragments of 649, 440 and 31bp were detected in propositus' brother, revealing a homozygotes mutation in b5R gene. Fragments of 649,471,440 and 31 bp were detected in his parents, revealing a heterozygous mutation in b5R gene. Conclusion Missense mutation (CTC-CCC)at codon 72 of b5R gene can be detected by using PCR technique, combined with Apa I endonuclease restriction analysis. PCR-RFLP is a simple, rapid and efficient method in detection of heterozygote carriers among human population.

关 键 词：遗传性高铁血红蛋白血症 细胞色素b5还原酶聚合酶链反应 限制性片段长度多态性 基因突变 

分 类 号：R596[医药卫生—内科学]