Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM

Shin Yu Lin, Yi Ning Su, Chia Cheng Hung, Woei Tsay, Shyh Shin Chiou, Chieh Ting Chang, Hong Nerng Ho, Chien Nan Lee

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Background: 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. Methods: 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. Results: 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. Conclusion: 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.

Original languageEnglish
Article number53
JournalBMC Medical Genetics
Volume9
DOIs
Publication statusPublished - Jun 20 2008
Externally publishedYes

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DNA Mutational Analysis
DNA Primers
Multiplex Polymerase Chain Reaction
Hemophilia A
Capillary Electrophoresis
Taiwan
Introns
Freezing
High Pressure Liquid Chromatography
Polymerase Chain Reaction
Mutation
Population
Hemorrhagic Disorders
Costs and Cost Analysis
Factor X
Factor VIII
Mutation Rate
Genes
Exons

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM. / Lin, Shin Yu; Su, Yi Ning; Hung, Chia Cheng; Tsay, Woei; Chiou, Shyh Shin; Chang, Chieh Ting; Ho, Hong Nerng; Lee, Chien Nan.

In: BMC Medical Genetics, Vol. 9, 53, 20.06.2008.

Research output: Contribution to journalArticle

Lin, Shin Yu ; Su, Yi Ning ; Hung, Chia Cheng ; Tsay, Woei ; Chiou, Shyh Shin ; Chang, Chieh Ting ; Ho, Hong Nerng ; Lee, Chien Nan. / Mutation spectrum of 122 hemophilia A families from Taiwanese population by LD-PCR, DHPLC, multiplex PCR and evaluating the clinical application of HRM. In: BMC Medical Genetics. 2008 ; Vol. 9.
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