Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM.
Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM.
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LD-PCR，DHPLC，Multiplex PCR和HRM的临床应用，来自台湾人口的122个血友病的突变光谱。
DOI：
10.1186/1471-2350-9-53
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发表时间：
2008-06-20
影响因子：
--
通讯作者：
Lee, Chien-Nan
中科院分区：
文献类型：
--
作者：
Lin, Shin-Yu;Su, Yi-Ning;Hung, Chia-Cheng;Tsay, Woei;Chiou, Shyh-Shin;Chang, Chieh-Ting;Ho, Hong-Nerng;Lee, Chien-Nan
A 型血友病是最常见和最严重的遗传性出血性疾病。它是由 F8 基因突变引起的，该突变导致因子 VIII 蛋白缺乏或功能障碍，因子 VIII 蛋白是因子 X 激活复合物中的重要辅助因子。我们采用长距离聚合酶链式反应和变性高效液相色谱法对F8基因进行突变扫描。我们设计了竞争性多重 PCR 来识别具有外显子缺失的载体。为了提高通量并最大限度地降低突变扫描的成本，我们还评估了一种新的突变扫描技术，即高分辨率熔解分析（HRM），作为替代筛选方法。我们展示了对 122 个台湾 A 型血友病家族进行详细筛查的结果，并报告了 29 个新突变。鉴定出1个全外显子缺失家族，多重PCR成功识别该携带者。通过 HRM，可以轻松识别 28 例中的 25 例 (89%) 和 15 例中的 15 例 (100%) 携带者的不同熔解曲线模式。灵敏度为 93% (40/43)。本研究中血友病A总体突变检出率为100%。我们提出了甲型血友病基因诊断的诊断策略。我们认为 HRM 是一种强大的筛查工具，可为我们提供更具成本效益的 A 型血友病突变鉴定方案。
Hemophilia A represents the most common and severe inherited hemorrhagic disorder. It is caused by mutations in the F8 gene, which leads to a deficiency or dysfunctional factor VIII protein, an essential cofactor in the factor X activation complex. We used long-distance polymerase chain reaction and denaturing high performance liquid chromatography for mutation scanning of the F8 gene. We designed the competitive multiplex PCR to identify the carrier with exonal deletions. In order to facilitate throughput and minimize the cost of mutation scanning, we also evaluated a new mutation scanning technique, high resolution melting analysis (HRM), as an alternative screening method. We presented the results of detailed screening of 122 Taiwanese families with hemophilia A and reported twenty-nine novel mutations. There was one family identified with whole exons deletion, and the carriers were successfully recognized by multiplex PCR. By HRM, the different melting curve patterns were easily identified in 25 out of 28 cases (89%) and 15 out of 15 (100%) carriers. The sensitivity was 93 % (40/43). The overall mutation detection rate of hemophilia A was 100% in this study. We proposed a diagnostic strategy for hemophilia A genetic diagnosis. We consider HRM as a powerful screening tool that would provide us with a more cost-effective protocol for hemophilia A mutation identification.
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影响因子：
4
作者：
COLEMAN, R;GENET, SA;WILKIE, AOM
通讯作者：
WILKIE, AOM
影响因子：
9.3
作者：
Hung, CC;Su, YN;Lee, CN
通讯作者：
Lee, CN
影响因子：
2.9
作者：
Hung, Chia-Cheng;Su, Yi-Ning;Hsu, Su-Ming
通讯作者：
Hsu, Su-Ming
影响因子：
30.8
作者：
LAKICH, D;KAZAZIAN, HH;GITSCHIER, J
通讯作者：
GITSCHIER, J
影响因子：
20.3
作者：
Bagnall, RD;Waseem, N;Giannelli, F
通讯作者：
Giannelli, F