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目的：检测一例重型血友病Ａ患者（ＳＨ９）的基因突变。方法：用ＰＣＲ、变性梯度凝胶电泳（ＤＧＧＥ）和ＤＮＡ测序检测因子Ⅷ基因突变。先用Ｓｏｕｔｈｅｒｎｂｌｏｔｉｎｇ排除内含子２２倒位，然后应用ＰＣＲ对凝血因子Ⅷ基因进行扩增。扩增范围包括所有外显子及其侧翼内含子序列。结果：片段１４２在ＤＧＧＥ中泳动异常。ＤＮＡ测序证实Ｃ２５３５Ａ导致Ｂ区错义突变８２６Ａｓｐ（ＧＡＣ）→Ｇｌｕ（ＧＡＡ）。结论：该突变可能是导致重型血友病Ａ的原因，但有待进一步研究证实。
Objective: To detect a gene mutation in a patient with severe hemophilia A (SH9). Methods: Factor Ⅷ gene mutation was detected by PCR, denaturing gradient gel electrophoresis (DGGE) and DNA sequencing. Southern blotting was used to exclude inversions of the intron 22 and then the factor VIII gene was amplified by PCR. The amplification range includes all exons and their flanking intron sequences. Results: Fragment 142 migrated abnormally in DGGE. DNA sequencing confirmed that C2535A caused a missense mutation in B region 826Asp (GAC) → Glu (GAA). Conclusion: This mutation may be the cause of severe hemophilia A, but it needs further study to confirm.