重型血友病甲基因诊断新技术的研究及其应用

Study on Developing New Technology of Diagnosis for Severe Hemophilia A and Its Application

运用长距离ＤＮＡ扩增技术（ＬＤ－ＰＣＲ）成功地检测出凝血八因子ＦⅧ基因倒位缺陷患者，并检出该倒位的女性携带者。在建立并完善上述技术之后，对５３份ＨＡ患者及家系成员的ＤＮＡ标本进行了双盲试验，检出ＦⅧ倒位者２３份，野生型２７份，携带者３份。结果与用Ｓｏｕｔｈｅｒｎ杂交技术检测的结果完全一致，说明运用本ＬＤ－ＰＣＲ技术检测ＦⅧ基因倒位的灵敏度与准确率均达到百分之百。运用这一新技术，完成了５个重型血友病甲家系的基因诊断。共检出３例基因倒位者，４位女性倒位基因的携带者。该技术较Ｓｏｕｔｈ－ｅｒｎ印迹杂交法简便、快速、高效直观、灵敏准确，便于推广应用，达到国际先进水平。

To detect Factor Ⅷ(FⅧ) gene inversion leading to severe Hemophilia A and carrier, a new technique based on the Long Distance Polymerase Chain Reaction(LD PCR) was developed to replace the Southern Blot method. The blind analyses of 53 DNA samples from HA patients and their families carried out by the LD PCR and Southern Blot respectively, and the same results were obtained: 23 cases of inversion, 27 cases of wild type, and 3 carriers. The sensitivity and specificity of the LD PCR are both 100%. Three inversion hemizygous and 4 female carriers were identified from 5 HA families by the LD PCR technology. Compared with the Southern Blot, the technique is simple, rapid, inexpensive, more sensitive and accurate.