The Human Leukocyte Antigen (HLA) system is a family of 21 genes found on chromosome 6. These genes exhibit high levels of polymorphism and more than 12,500 HLA alleles have now been characterized in the IPD-IMGT/HLA database.
This high level of polymorphism enables the immune system to distinguish between host cells, non-host cells, infected cells and dysfunctional cells. Certain HLA alleles have also been associated with some types of cancer and several autoimmune disorders including rheumatoid arthritis, type 1 diabetes and inflammatory bowel diseases. HLA typing is therefore a valuable application used for donor–recipient HLA matching to reduce the risk of transplant rejection and as a diagnostic tool for establishing the risk of some HLA-associated disorders.
Scientists from Beckman Coulter have recently described automation of the Illumina TruSight® HLA Sequencing Panel on the Beckman Coulter Biomek® FXP HLA SP, which is an application specific configuration of the Biomek® FXP Dual Multi-96 Span-8 automated liquid handler. This sequencing panel can be used to prepare genomic DNA libraries that enable high resolution, unambiguous HLA sequencing with a single assay.
Once the HLA loci are amplified, uniquely indexed libraries are produced for each locus in all the genomic DNA samples and then sequenced using the MiSeq® system. The standardization of library preparation using Biomek FXP TruSight HLA automation provides significant time savings over manual preparation, with up to 192 libraries being created from 24 genomic DNA samples in around 11 hours. This method reduces the need for operator interaction, which lowers the incidence of costly pipetting errors and provides more uniform sample processing.
The libraries generated can be loaded directly onto the MiSeq® Reagent Cartridge and sophisticated data handling accelerates preparation of the required Sample Sheet. To provide further savings on time and consumables, an integrated Biometra TRobot thermocycler is available for on-deck incubations, individual Span-8 probes perform enzyme and reagent transfers and the steps involved in sample clean-ups are carried out by a multi-channel 96 pipetting head.
Analysis of the sample data generated using this automation method shows the data is highly reproducible compared with data generated manually. Automated runs have also been devoid of contaminating sequences from neighboring wells, indicating that no cross-contamination occurs between samples.
The Biomek FXP TruSight HLA automation method streamlines the workflow of the TruSight HLA Sequencing Panel and generates accurate and highly reproducible HLA typing results.
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About Beckman Coulter
Beckman Coulter develops, manufactures and markets products that simplify, automate and innovate complex biomedical tests. More than a quarter of a million Beckman Coulter instruments operate in laboratories around the world, supplying critical information for improving patient health and reducing the cost of care.
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