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目的 探索一种更简便 ,更特异的方法 ,用于血友病甲的基因诊断及其家系的遗传咨询。方法 应用聚合酶链反应 (PCR)和双链异源泳动分析 (HMA)对我国 10个血友病甲患儿及其家系成员中FⅧ基因内 13内含子二核苷酸重复序列 (CA)n多态性和等位基因进行分析。结果 10个家系中 ,男性 2 0例 ,女性 10例 ,共 4 0条X染色体 ,经PCR扩增、琼脂糖电泳均可显示出 14 0～ 15 0bpDNA条带 ,男性呈现单一的DNA条带 ,女性则根据等位基因的不同 ,出现两条DNA条带 (杂合子 )或者一条DNA条带 (纯合子 )。结论 二核苷酸重复序列广泛分布于中国人群FⅧ基因中 ,重复序列的多态性作为第二代遗传标志 ,结合等位基因分析和HMA杂交电泳泳动分析 ,可明确鉴别出血友病甲基因的携带者 ,可用于血友病甲的基因诊断和遗传咨询
Objective To explore a simpler and more specific method for the genetic diagnosis of hemophilia A and the genetic counseling of its pedigree. Methods Polymerase chain reaction (PCR) and double-stranded heterologous electrophoresis (HMA) were used to detect the nucleotide sequence of intron 13 dinucleotide repeats of FⅧ gene in 10 children with hemophilia A and their pedigrees (CA ) n polymorphisms and alleles were analyzed. Results In 10 pedigrees, there were 20 males and 10 females with a total of 40 X chromosomes. The 14 0 ~ 150 bp DNA bands were detected by PCR amplification and agarose gel electrophoresis. The DNA bands were single in males. Depending on the allele, women show two DNA bands (heterozygotes) or one DNA band (homozygotes). CONCLUSION: The dinucleotide repeats are widely distributed in the FⅧ gene of Chinese population. The polymorphism of the repeats is the second generation genetic marker. Combined with allele analysis and HMA hybridization electrophoresis, it is possible to clearly identify the hemophilia A gene Carriers, can be used for genetic diagnosis and genetic counseling Hemophilia A.