title: High-Throughput Sequencing of the Major Histocompatibility Complex following Targeted Sequence Capture.
abstract: The Human Major Histocompatibility Complex (MHC) is a highly polymorphic region full of immunoregulatory genes. The MHC codes for the human leukocyte antigens (HLA), proteins that present on the cellular surface and that are involved in self-non-self recognition. For matching donors and recipients for organ and stem-cell transplants it is important to know an individual's HLA haplotype determinable in this region. Now, as next-generation sequencing (NGS) platforms mature and become more and more accepted as a standard method, NGS applications have spread from research laboratories to the clinic, where they provide valid genetic insights. Here, we describe a cost-effective microarray-based sequence capture, enrichment, and NGS sequencing approach to characterize MHC haplotypes. Using this approach, ~4 MB of MHC sequence for four DNA samples (donor, recipient and the parents of the recipient) were sequenced in parallel in one NGS instrument run. We complemented this approach using microarray-based genome-wide SNP analysis. Taken together, the use of recently developed tools and protocols for sequence capture and massively parallel sequencing allows for detailed MHC analysis and donor-recipient matching.
authors.affiliation: Red Cross Transfusion Service of Upper Austria, Krankenhausstr. 7, 4020, Linz, Austria. Johannes.Proell@o.roteskreuz.at.
authors.forename: Johannes
authors.initials: J
authors.lastname: Pröll
authors.affiliation: Red Cross Transfusion Service of Upper Austria, Krankenhausstr. 7, 4020, Linz, Austria.
authors.forename: Carina
authors.initials: C
authors.lastname: Fischer
authors.affiliation: Center for Medical Research, Graz, Austria.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Michelitsch
authors.affiliation: Red Cross Transfusion Service of Upper Austria, Krankenhausstr. 7, 4020, Linz, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Danzer
authors.affiliation: Red Cross Transfusion Service of Upper Austria, Krankenhausstr. 7, 4020, Linz, Austria.
authors.forename: Norbert
authors.initials: N
authors.lastname: Niklas
mesh_terms: D005838:Genotype; D006680:HLA Antigens / Q000235:genetics; D006239:Haplotypes / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008285:Major Histocompatibility Complex / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D006680:HLA Antigens
keywords: Major histocompatibility complex; Next-generation sequencing; Targeted sequence capture