The use of high resolution melting analysis to detect Fabry mutations in heterozygous females via dry bloodspots
| Autor: | Ching-Yuang Lin, Chang-Long Tai, Shu Min Kao, Chiang-Chuan Chiang, Hsiao-Chi Yu, Y.-H. Huang, Chia-Feng Yang, Jan-Gowth Chang, Fang-Chih Tsai, Jeng-Hung Suen, Hung Chiang, Tina Jui-Ting Wu, Hong-Duo Chen, Mei-Ying Liu, Dau-Ming Niu |
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| Rok vydání: | 2011 |
| Předmět: |
Male
Heterozygote Clinical Biochemistry DNA Mutational Analysis Biology medicine.disease_cause Nucleic Acid Denaturation Biochemistry High Resolution Melt law.invention law medicine Intronic Mutation Humans Transition Temperature Polymerase chain reaction Genetics Mutation Newborn screening Biochemistry (medical) Genetic disorder Infant Newborn General Medicine Exons medicine.disease Fabry disease Fabry Disease Female Dried Blood Spot Testing Asymptomatic carrier |
| Zdroj: | Clinica chimica acta; international journal of clinical chemistry. 413(3-4) |
| ISSN: | 1873-3492 |
| Popis: | Background As an X-linked genetic disorder, Fabry disease was first thought to affect males only, and females were generally considered to be asymptomatic carriers. However, recent research suggests that female carriers of Fabry disease may still develop vital organ damage causing severe morbidity and mortality. In the previous newborn screening, from 299,007 newborns, we identified a total of 20 different Fabry mutations and 121 newborns with Fabry mutations. However, we found that most female carriers are not detected by enzyme assays. Methods A streamlined method for high resolution melting (HRM) analysis was designed to screen for GLA gene mutations using a same PCR and melting program. Primer sets were designed to cover the 7 exons and the Chinese common intronic mutation, IVS4+919G>A of GLA gene. Results The HRM analysis was successful in identifying heterozygous and hemizygous patients with the 20 surveyed mutations. We were also successful in using this method to test dry blood spots of newborns afflicted with Fabry mutations without having to determine DNA concentration before PCR amplification. Conclusion The results of this study show that HRM could be a reliable and sensitive method for use in the rapid screening of females for GLA mutations. |
| Databáze: | OpenAIRE |
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