Chapter title |
High-Throughput Sequencing of the Major Histocompatibility Complex following Targeted Sequence Capture
|
---|---|
Chapter number | 5 |
Book title |
Haplotyping
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6750-6_5 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6748-3, 978-1-4939-6750-6
|
Authors |
Johannes Pröll, Carina Fischer, Gabriele Michelitsch, Martin Danzer, Norbert Niklas |
Editors |
Irene Tiemann-Boege, Andrea Betancourt |
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. |
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