兰州大学机构库 >学院待认领
中国西北地区血友病B的基因诊断
Alternative TitleThe genetic diagnosis of hemophilia B in northwest of china
曾蕾
Thesis Advisor李燕平
2011-05-16
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword血友病B 基因型-表型相互关系 STR位点 直接测序 基因突变
Abstract研究背景: 血友病B(Hemophilia B HB)是由于凝血因子Ⅸ(factorⅨ,FIX)的基因突变,导致了血浆FIX数量的缺乏或者质量的缺陷,从而引起的X连锁隐性遗传性出血性疾病。本文为了研究西北地区HB患者的FⅨ基因的突变谱的特征,建立最优化的分子诊断策略,找出基因型-表型的相互关系,为HB的早期准确性诊断、治疗和预防提供科学的依据。 方法: 1. 取4个无亲缘关系的HB家系(共23人)成员、3名无亲缘关系的散发HB患者血清样本,进行DNA的提取。 2. 通过常规检测患者的PT、APTT和FⅨ:C来进行筛查及确诊。 3. 利用PCR方法对4个家系成员扩增6个STR位点片段,检测各个成员的6个位点的多态性,进行4个家系的遗传连锁分析。 4. 利用PCR方法对3位HB患者的8个FⅨ基因外显子序列进行扩增,用直接测序法对扩增产物进行测序,其结果用Chromas与正常序列对比,找出突变的基因。 结果 1. 选择在国人中具有一定多态性频率的FⅨ基因内外的6个STR位点,对4个血友病B家系共23人进行基因连锁分析, 其中7个HB患者通过全部检出,另外还检出6个携带者。 2. 用直接测序方法对3名无家族史的散发HB患者检测到3个不同的突变,(突变检测率达到了100%)1号患者有外显子2点突变(A→C);2号患者有外显子8点突变(G→A);3号有外显子2点突变(C/T型)。 结论 1. 在所选的HB患者的基因突变中未发现突变热点。 2. 突变类型和基因缺失的严重程度的基因型-表型不相关。 3. 联合6个STR位点对西北地区HB家系患者的遗传分析是有效的间接诊断方法。 4. 直接测序8个外显子和侧翼序列被认为是血友病B患者特别是无家族史的散发病例的诊断金标准。
Other AbstractBackground Hemophilia B(HB) is due to coagulation factor Ⅸ (factor Ⅸ, FIX) gene mutation, resulting in a lack of volume of plasma FIX or qualitative defects, causing X-linked recessive hereditary bleeding disorder. The article is to study the mutation spectrum of F9 in the Northwest patients with HB , to establish the most optimized molecular diagnostic strategies, to identify genotype - phenotype relationship for HB and to seek evidence for accurate early diagnosis, treatment and prevention . Methods 1. Four in northwest HB unrelated families (23) members, three in northwest HB unrelated patients were taked extracted DNA. 2. PT , APTTand F Ⅸ: C by routine testing of patients for phenotype diagnosis 3.Four family members on six STR loci fragments are amplified By PCR,each member of the six detected polymorphism, the four families of genetic linkage analysis. 4. Three HB patients of FⅨwith exon sequences were amplified by PCR method, amplified products were sequenced by direct sequencing , and the results compared with the normal sequence for Chromas ,find out mutations in the gene. Results 1. Joint bring 6 STR sites high polymorphic information in China , on the four families with hemophilia B (23), 7 HB carriers, for genetic linkage analysis, 7 HB Patients were all detected, in addition to the detection 6 A carriers. 2. A total of 3 unique mutations were detected By direct sequencing in three HB patients with no family history, (mutation detection rate100%),the point mutation of exon 2 was found in patient No 1(A→C) ; the point mutation of exon8 existed in patient No 2(G→A); the point mutation of exon2 was detected in patient No 3 (C/T). Conclusion 1. HB patients in test with no hotspots for mutations. 2. The genotype–phenotype correlations between the type of mutation and the severity of factor deficiency were not consistent. 3. Joint 6 STR loci in HB patients of Northwest for Genetic analysis of families is an effective method of indirect diagnosis 4. Direct sequencing of all exons and flanking sequences are considered the gold standard diagnosis for the sporadic cases, especially in patients without a family history ones.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222210
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
曾蕾. 中国西北地区血友病B的基因诊断[D]. 兰州. 兰州大学,2011.
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