Department of Haemostasis, National Institute of Immunohaematology (ICMR), KEM Hospital Campus, Parel, Mumbai 400012, India.
Clin Chim Acta. 2010 Dec 14;411(23-24):2004-8. doi: 10.1016/j.cca.2010.08.026. Epub 2010 Aug 26.
The identification of pathogenic mutations in haemophilia A (HA) patients is important as a basis for genetic diagnosis and also for the assessment of clinical manifestations.
We analyzed 36 inversion negative congenital HA cases (28 unrelated and 8 familial) by multiplex PCR and the conformation sensitive gel electrophoresis (CSGE) technique, followed by DNA sequencing. The pathogenicity of each of these mutations was assessed using various prediction software.
We found 17 missense, 5 deletions, 3 insertions, and 2 nonsense mutations, out of which 16 were recurrent and 11 novel. All novel substitution mutations were found to be deleterious using the prediction softwares. We also encountered a double mutation (1 novel and 1 hot-spot mutation) in a family with a strong family history. A missense mutation in heterozygous state was also detected in a female bleeder with very low factor VIII levels, probably due to extreme lyonization.
High heterogeneity in mutational profile has been observed in the present study. The outcome of this study would enable us to give an accurate diagnosis in all affected families by direct mutation analysis.
在甲型血友病（HA）患者中，确定致病突变非常重要，这不仅是遗传诊断的基础，也是评估临床表现的基础。
我们通过多重 PCR 和构象敏感凝胶电泳（CSGE）技术分析了 36 例反转阴性先天性 HA 病例（28 例非相关和 8 例家族性），然后进行 DNA 测序。使用各种预测软件评估了每个突变的致病性。
我们发现了 17 个错义突变、5 个缺失突变、3 个插入突变和 2 个无义突变，其中 16 个是重复的，11 个是新的。所有新的替代突变都被预测软件认为是有害的。我们还在一个有强烈家族史的家庭中发现了一个双重突变（1 个新突变和 1 个热点突变）。在一个因子 VIII 水平极低的女性出血者中，还检测到了杂合状态的错义突变，可能是由于极端里昂化所致。
本研究观察到突变谱的高度异质性。本研究的结果将使我们能够通过直接突变分析对所有受影响的家庭做出准确的诊断。
