Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Stabentheiner, S; Danzer, M; Niklas, N; Atzmüller, S; Pröll, J; Hackl, C; Polin, H; Hofer, K; Gabriel, C.
Overcoming methodical limits of standard RHD genotyping by next-generation sequencing.
Vox Sang. 2011; 100(4): 381-388.
Doi: 10.1111/j.1423-0410.2010.01444.x
Web of Science
PubMed
FullText
FullText_MUG
- Co-Autor*innen der Med Uni Graz
- GABRIEL Christian
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- Abstract:
- Molecular variations of the RHD gene may result in the reduced expression of the D antigen and altered Rh phenotypes. In many occasions, they cannot be typed reliably by standard serological methods. Sequence-based typing is the gold standard to determine rare and unknown RHD genotypes. For this pilot study, sequence-based typing by standard Sanger sequencing was compared to a newly established next-generation sequencing approach based on pyrosequencing. Twenty-six DNA samples were selected after primary serological testing exhibiting a weak reaction in Rh phenotype. Parallel sequence analysis of the complete coding sequence including adjacent intronic sequences allowed a comparison of the methodical potency in mutation detection of Sanger with next-generation sequencing. Sanger sequencing revealed 39 RHD polymorphisms in 21 of 26 samples in the RHD coding region, while pyrosequencing detected all but two alterations resulting in a concordance rate of 94·9% and clearly revealed a heterozygous compound mutation in one sample with RHDψ and Weak D type 4 alleles. The resolution of cis/trans linkage of polymorphisms and exact characterization of a 37 bp duplication was achieved by next-generation sequencing. Our data suggest that next-generation sequencing offers a new development for high-throughput and clonal sequencing for molecular RHD genotyping. However, further attempts in the methodical set-up have to be undertaken prior to validation and introduction as a routine service. © 2010 The Author(s). Vox Sanguinis © 2010 International Society of Blood Transfusion.
- Find related publications in this database (using NLM MeSH Indexing)
- Blood Grouping and Crossmatching - methods Blood Grouping and Crossmatching - standards
- Female -
- Humans -
- Male -
- Polymorphism, Genetic -
- Rh-Hr Blood-Group System - genetics
- Sequence Analysis, DNA - methods Sequence Analysis, DNA - standards
- Find related publications in this database (Keywords)
- genotyping
- next-generation sequencing
- RHD
- sequence-based typing