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目的对1例遗传性凝血因子ⅫⅠ(FⅫⅠ)缺陷症患者及其家系成员 FⅫⅠA基因[F(13)A]进行分析,探讨其分子致病机制。方法尿素溶解法定性检测 FⅫⅠ活性,抽提外周血基因组 DNA,PCR 扩增 FⅫⅠA基因的15个外显子及其侧翼序列,PCR 产物纯化后直接基因测序,并对家系成员F(13)基因相应的突变序列进行检测。结果先证者 FⅫⅠ定性试验阳性。基因测序发现先证者F(13)A存在纯合缺失,外显子10自127067位起缺失33个核苷酸(127067del33,GI:AF418272),导致阅读框内缺失11个氨基酸(406Met-416Ala),产生了由720个氨基酸残基组成的截短型 FⅫⅠA蛋白。其父母 FⅫⅠ定性试验为阴性,均显示为该序列的杂合缺失突变。结论先证者为遗传性 FⅫⅠ缺陷症患者,由 F(13)A 外显子10缺失突变所致。该突变为国际首次报道。
OBJECTIVE: To investigate the molecular pathogenesis of FⅫⅠA gene [F (13) A] in patients with hereditary clotting factor ⅫⅠ (FⅫⅠ) deficiency and their family members. Methods The FⅫⅠactivities were qualitatively determined by urea dissolution. Genomic DNA was extracted from peripheral blood. Fifteen exons and their flanking sequences of FⅫⅠA gene were amplified by PCR. The PCR products were directly sequenced after purification. The F (13) The mutation sequence was detected. Results proband F Ⅻ qualitative test was positive. Gene sequencing revealed homozygous deletion of proband F (13) A, loss of 33 nucleotides from 127067 (127067del33, GI: AF418272) in exon 10, resulting in the deletion of 11 amino acids in the reading frame (406 Met-416Ala ), A truncated FⅫIA protein consisting of 720 amino acid residues was generated. Their parents F Ⅻ qualitative test was negative, showed that the sequence of heterozygous deletion mutation. Conclusion The probands are patients with hereditary F Ⅻ I deficiency and are caused by the deletion of F (13) A exon 10 mutations. The mutation was first reported in the world.