论文部分内容阅读
目的探讨血友病A(HA)患者及携带者的基因诊断方法及诊断率。方法对20例HA患者及家系成员采用长距离PCR扩增(LD-PCR)技术、限制性内切酶酶切位点连锁分析(BclⅠPCR/RFLP)、可变串联重复序列多态性分析(St14 VNTR/PCR);并对FⅧ基因18号内含子的BclⅠ区段、19号内含子的HindⅢ相应区段进行DNA测序。结果检出22号内含子倒位携带者1例和倒位者6例;BclⅠPCR/RFLP酶切位点突变患者3例,携带者5例;St14 VNTR家系的连锁分析,发现携带者6例,患者5例;联合诊断家系总阳性率为90.0%;DNA测序两个区段突变位点分析,共发现5种突变。结论采用联合基因诊断方法,对FⅧ基因的3个位点进行联合基因诊断,可提高HA的诊断率及准确率;用PCR-DNA测序法可发现5种突变,并且这些突变位点在国内报道较少,22例患者突变检出率达50.0%。
Objective To investigate the gene diagnosis and diagnosis rate of hemophilia A (HA) patients and carriers. Methods Twenty cases of HA patients and their pedigree were analyzed by long distance PCR (LD-PCR), restriction endonuclease analysis (BclⅠPCR / RFLP), variable tandem repeat polymorphism analysis (St14 VNTR / PCR). The BclI segment of FⅧ gene intron 18 and the corresponding segment of HindⅢ in intron 19 were sequenced. Results 1 intron 22 intron and 6 inversions were detected in the intron. There were 3 patients with BclⅠPCR / RFLP restriction site mutation and 5 carriers. St14 VNTR pedigree linkage analysis found 6 carriers , 5 cases of patients; combined diagnosis of pedigrees total positive rate was 90.0%; DNA sequencing mutations in the two segments, found a total of 5 mutations. Conclusion Combined gene diagnosis method can improve the diagnosis rate and accuracy of HA in three loci of FⅧ gene. Five mutations were found by PCR-DNA sequencing, and these mutations were reported in China Less, 22 patients with mutation detection rate of 50.0%.