Mutation Spectrum of Six Genes in Chinese Phenylketonuria Patients Obtained through Next-Generation Sequencing
| Autor: | Lin Lin, Jing Hao, Ying Gu, Cen Zhong, Zhigang Yang, Cui Shujian, Yang Guanghui, Jian Huang, Li Yu, Kangmo Lu, Jiabao Peng |
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| Rok vydání: | 2014 |
| Předmět: |
China
Phenylketonurias DNA Mutational Analysis Nonsense mutation lcsh:Medicine Biology DNA sequencing law.invention Autosomal Recessive Diseases Genomic Medicine Asian People INDEL Mutation law Genetics Medicine and Health Sciences Phenylketonuria Humans Genetic Testing Genome Sequencing Molecular Biology Techniques Sequencing Techniques lcsh:Science Molecular Biology Gene Polymerase chain reaction Clinical Genetics Autosomal Recessive Traits Multidisciplinary Point mutation lcsh:R Biology and Life Sciences Computational Biology High-Throughput Nucleotide Sequencing Human Genetics Genomics Biopterin Pedigree Mutation Mutation (genetic algorithm) lcsh:Q Genetic Dominance Multiplex Polymerase Chain Reaction Research Article |
| Zdroj: | PLoS ONE, Vol 9, Iss 4, p e94100 (2014) PLoS ONE |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0094100 |
| Popis: | BACKGROUND: The identification of gene variants plays an important role in the diagnosis of genetic diseases. METHODOLOGY/PRINCIPAL FINDINGS: To develop a rapid method for the diagnosis of phenylketonuria (PKU) and tetrahydrobiopterin (BH4) deficiency, we designed a multiplex, PCR-based primer panel to amplify all the exons and flanking regions (50 bp average) of six PKU-associated genes (PAH, PTS, GCH1, QDPR, PCBD1 and GFRP). The Ion Torrent Personal Genome Machine (PGM) System was used to detect mutations in all the exons of these six genes. We tested 93 DNA samples from blood specimens from 35 patients and their parents (32 families) and 26 healthy adults. Using strict bioinformatic criteria, this sequencing data provided, on average, 99.14% coverage of the 39 exons at more than 70-fold mean depth of coverage. We found 23 previously documented variants in the PAH gene and six novel mutations in the PAH and PTS genes. A detailed analysis of the mutation spectrum of these patients is described in this study. CONCLUSIONS/SIGNIFICANCE: These results were confirmed by Sanger sequencing. In conclusion, benchtop next-generation sequencing technology can be used to detect mutations in monogenic diseases and can detect both point mutations and indels with high sensitivity, fidelity and throughput at a lower cost than conventional methods in clinical applications. |
| Databáze: | OpenAIRE |
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