Bioinformatic integration of biomechanics makes HLA sequencing universally applicable

Sequencing provides the most reliable and accurate information of the sequence of a gene and is therefore of particular interest for fully characterizing the genetic complexity and allelic diversity of human leukocyte antigens (HLA) genes. This complexity makes sequencing the method of choice for HLA low- and high-resolution typing. However, sequencing is considered a complex process, which requires the management of an overwhelming amount of data. However, only sequencing can be used as a sole typing technique without the need of supplementation by other typing techniques; it offers additional advantages by providing results in electronic formats and by being fully automatable. This makes sequencing the best candidate for bioinformatic and biomechanic integration to maintain the benefits and discard the drawbacks. We have, therefore, developed robotic platforms and a bioinformatic tool allowing complete documentation, interpretation and processing of all amplification and sequencing steps as well as the full integration of all instruments involved. It was shown that this integrative approach allows sequencing to run fully automated for low- and high-resolution typing on each throughput level desired. This integration of robotics and software secures the traceability of each specimen at each step of the process and minimizes the risk of contamination and sample mix-up.