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目的最大限度提高血友病A(hemophiliaA,HA)患者及家系成员的基因诊断、携带者检出。方法对22例HA患者和两个家系的成员14人首先采用长距离DNA扩增(LD-PCR)技术,直接检测是否为基因倒位及其携带者;对于非倒位的HA家系依次采用FⅧ基因内的位点BclⅠ(RFLP)、内含子13和22中的STR、FⅧ基因外的DXS52(ST14)位点的多态性进行遗传连锁分析。结果22例HA中12例为重型,8例为中型,2例轻型;12例重型中6例为内含子22倒位,占50%,用该基因信息为一家系2名女性进行了检测均为携带者;联合应用间接遗传连锁分析,对2个家系进行检测,家系C中有1名女性为携带者,家系D中2名女性确定为携带者,1名为正常人。结论联合应用四种分子生物学技术,几乎可以为所有有家族史的HA家系作出基因诊断和携带者检出。
Objective To maximize the genetic diagnosis and carrier detection of hemophilia A (HA) patients and their family members. Methods Twenty-two HA patients and 14 individuals from two pedigrees were investigated by direct DNA amplification (LD-PCR) technique to determine whether they were gene inversion and their carriers. For non-inverted HA families, FⅧ Genetic polymorphism of the locus BclI (RFLP), the STRs in intron 13 and 22, and the DXS52 (ST14) locus outside of the FⅧ gene were analyzed by genetic linkage analysis. Results Of the 22 cases, HA was found in 12 cases of heavy type, 8 cases of moderate type and 2 cases of light type. In 6 cases of 12 cases of heavy type, intron 22 was inverted, accounting for 50% All of them were carriers. Using indirect genetic linkage analysis, two families were tested. One female was a carrier in family C and two women in family D were carriers and one was normal. Conclusions The combined use of four molecular biological techniques allows genetic diagnosis and carrier detection in almost all HA families with family history.