Next generation sequencing of SNPs for non-invasive prenatal diagnosis: challenges and feasibility as illustrated by an application to β-thalassaemia
| Autor: | Marina Kleanthous, Mirjam C G N van den Hout, Wilfred F. J. van IJcken, Christel E M Kockx, Loukas Kythreotis, Eleni Kalogirou, Thessalia Papasavva, Petros Kountouris, Frank Grosveld |
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| Přispěvatelé: | Cell biology |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Population
Locus (genetics) Single-nucleotide polymorphism Prenatal diagnosis Biology Polymorphism Single Nucleotide Article Loss of heterozygosity 03 medical and health sciences 0302 clinical medicine Pregnancy Prenatal Diagnosis Genetics medicine Humans Allele education Genetics (clinical) Alleles 030304 developmental biology 0303 health sciences education.field_of_study 030219 obstetrics & reproductive medicine Haplotype beta-Thalassemia Beta thalassemia High-Throughput Nucleotide Sequencing medicine.disease 3. Good health Haplotypes Mutation Female |
| Zdroj: | European Journal of Human Genetics European Journal of Human Genetics, 21(12), 1403-1410. Nature Publishing Group |
| ISSN: | 1018-4813 |
| Popis: | beta-Thalassaemia is one of the most common autosomal recessive single-gene disorder worldwide, with a carrier frequency of 12% in Cyprus. Prenatal tests for at risk pregnancies use invasive methods and development of a non-invasive prenatal diagnostic (NIPD) method is of paramount importance to prevent unnecessary risks inherent to invasive methods. Here, we describe such a method by assessing a modified version of next generation sequencing (NGS) using the Illumina platform, called 'targeted sequencing', based on the detection of paternally inherited fetal alleles in maternal plasma. We selected four single-nucleotide polymorphisms (SNPs) located in the beta-globin locus with a high degree of heterozygosity in the Cypriot population. Spiked genomic samples were used to determine the specificity of the platform. We could detect the minor alleles in the expected ratio, showing the specificity of the platform. We then developed a multiplexed format for the selected SNPs and analysed ten maternal plasma samples from pregnancies at risk. The presence or absence of the paternal mutant allele was correctly determined in 27 out of 34 samples analysed. With haplotype analysis, NIPD was possible on eight out of ten families. This is the first study carried out for the NIPD of beta-thalassaemia using targeted NGS and haplotype analysis. Preliminary results show that NGS is effective in detecting paternally inherited alleles in the maternal plasma. |
| Databáze: | OpenAIRE |
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