HLA領域のシーケンスを行うことにより、さまざまな免疫疾患に関する重要な知見を得ることができます。従来法によるヒト白血球型抗原(HLA)タイピングで高解像度の結果を得るためには、アッセイを繰り返し行う必要があり、複数のシステムや解析プログラムの運用が欠かせません。
次世代シーケンサー(NGS)を用いたHLAタイピングなら、1台のシステム、1つの解析プログラムによるフェーズされた明瞭なHLAタイピング結果が1回のアッセイで得られます。
The HLA region is the most densely polymorphic region of the genome. Sequencing HLA genes used to be notoriously difficult due to high levels of sequence homology between HLA genes and pseudogenes, dense variability, and poor characterization of the thousands of HLA alleles in the population.
NGS with Illumina sequencing by synthesis (SBS) chemistry overcomes these challenges, enabling simple, high-quality analysis of the key HLA genes. Highly accurate and robust Illumina sequencing by synthesis (SBS) chemistry is the most successful and widely adopted NGS technology worldwide.*
In principle, NGS is similar to Sanger (capillary electrophoresis-based) sequencing. The bases of a DNA fragment are identified sequentially from signals emitted as each fragment is resynthesized from a DNA template strand. NGS scales up this process; millions of reactions occur in a massively parallel fashion. This advance enables rapid sequencing of large stretches of DNA such as the HLA region.
Illumina NGS technology supports paired-end sequencing, a unique feature that is crucial for successful, unambiguous HLA typing. Sequencing the ends of the library DNA fragments generates high-quality base calls. The physical link between the 2 reads (originating from the same clonally amplified library DNA fragment) allows association of variants found in each read pair. The distance between the paired reads varies as a result of the random library fragment generation process, allowing the direct resolution of the phase of 2 variants.
RNA-Seq and HLA typing are increasing the power and efficiency of a target discovery platform.
Researchers query the genome with NGS and perform HLA genotyping to identify novel risk loci for immune-mediated disease.
Drawing upon the genetic diversity of Brazilians, researchers develop an HLA variation database.
Multiplexing enables large numbers of loci to be sequenced simultaneously during a single experiment. Individual “barcode” sequences are added to each locus so they can be differentiated during HLA sequencing data analysis.
TruSight HLA シーケンスパネル v2は、11のHLA遺伝子座の高解像度なタイピングが行える、HLA解析用に最適化された試薬およびソフトウェアを付属した、サンプルからレポートまでの包括的なソリューションです。その簡潔なワークフローでは、48時間未満のターンアラウンドタイムと4.5時間未満のハンズオンタイムでより一層の効率化が実現しています。
NGS and microarray solutions enable researchers to investigate potential autoimmune disease mechanisms, the immune repertoire, tumor immunogenicity, and the functional consequences of immune-related genetic variation.
*Data calculations on file. Illumina, Inc., 2015