High-throughput detection of common sequence variations of Fabry disease in Taiwan using DNA mass spectrometry
| Autor: | Dau-Ming Niu, Hsiang-Yu Lin, Cheng-Fang Li, Sheng-Hung Lee, Shih-Feng Tsai, Chien-Hsing Lin, Hao-Chuan Liu |
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| Rok vydání: | 2014 |
| Předmět: |
Male
Endocrinology Diabetes and Metabolism Population Taiwan Biology Biochemistry Polymerase Chain Reaction Mass Spectrometry symbols.namesake chemistry.chemical_compound Endocrinology Genetics medicine Humans education Dried blood Molecular Biology Genotyping Whole blood Sanger sequencing education.field_of_study Newborn screening Base Sequence High-Throughput Nucleotide Sequencing DNA medicine.disease Fabry disease Molecular biology chemistry Mutation symbols Fabry Disease Female |
| Zdroj: | Molecular genetics and metabolism. 111(4) |
| ISSN: | 1096-7206 |
| Popis: | Background In view of the therapeutic benefits resulting from early intervention for Fabry disease, our team has implemented an enzyme-based newborn screening in Taiwan since 2008. However, we found that most heterozygous females cannot be detected. To improve the screening efficiency, a more effective method for GLA gene genotyping is necessary. Methods As the suspected mutations are limited to only 29 different spots in Taiwanese, a panel of Sequenom iPLEX assay was designed for rapid screening of GLA variations. To determine the accuracy and sensitivity of this assay, previously diagnosed and undiagnosed DNA samples were analyzed by this genotyping assay and Sanger sequencing. In addition, DNA extracted from dried blood spots was also tested. Results Sequenom iPLEX assay is accurate and cost-effective, identifying the sequence variations, which were designated in the panel. It identified common GLA variants in DNA samples extracted from whole blood or dried blood spots with 100% accuracy and sensitivity. Conclusions Sequenom iPLEX assay is suitable for Fabry newborn screening when hotspot mutations and common variations are known in a well-studied population. In addition, this assay can also be applied for first-line determination of GLA variant sequences in suspected subjects of high-risk patients, or newborns. |
| Databáze: | OpenAIRE |
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