ASHI was also very educational for us and proved to be an excellent platform to identify the current trends in the HLA field. All the scientific presentations at the conference were outstanding, and provided us with a better understanding on what the HLA community is working on. We noticed that many are investigating the KIR domain, both in the fields of transplantation and disease association studies. Regarding molecular assays, next-generation sequencing-based technologies have been expanding on the market and catching up with PCR-based (SSO & SSR) methods, while Nanopore sequencing is a further development that seems to be gaining interest.

Among the posters we were pleased to read customers reporting on the benefits of using NGS and Omixon’s unique de novo assembly Consensus Genotyping algorithm over legacy methods (Plunkett et al, Binder et al, Gomes et al), and the protocols developed by Melista et al and Breeding et al both demonstrated how liquid handling automation significantly reduces hands-on time for Holotype HLA improving the accuracy and reproducibility of the assay.

Genetic investigations of the KIR gene cluster are of a continuous high interest among the community, both in the fields of transplantation and disease-association studies. Regarding the former, antibody-mediated rejection (AbMR) is the major barrier to long-term survival. Although its underlying mechanism remains to be understood, NK-cells seem to be the major effectors through their KIR domains. In fact, different KIR genotypes and gene motifs were shown to influence survival rates upon lung transplantation (Sun et al) and hematopoietic stem cell transplantation (Kurbegovic-Skaljic et al, Bao et al). Based on these observations a few laboratories have already adapted their donor selection procedure and included KIR genotyping results in their process (Sauter et al, Wagner et al, Lang et al). Significant contributions are also underway to extend the KIR genotype database given that the aformentioned labs have screened the loci in over 2 million samples since 2014. KIR-HLA ligand interactions, and KIR gene profiles also confer risk for several diseases such as multiple myeloma (Sun et al), myelogenous and lymphocytic leukemia (Akhter et al), nasopharyngeal carcinoma (Gao et al), pulmonary tuberculosis (Rajalingam), and rheumatoid arthritis (Ishitani et al).

We are rather interested to see whether more laboratories are going to apply routine KIR typing in the clinic, and how it will affect the development of genotyping products targeting the gene cluster.

With regard to cutting-edge technologies, a few presentations focused on nanopore sequencing. The MinION platform sequences long, single-stranded DNA as it passes through a nanopore and works by taking electrical current measurements. The technology needs to be improved since it has a relatively high error rate compared to current NGS methods. Among the posters, Clark et al performed single molecule sequencing of full length HLA amplicons using Omixon’s Holotype HLA kit. With an average read depth of over 200X per locus, their pipeline gave unambiguous genotyping results for each HLA locus that were also consistent with those generated from paired-end Illumina sequencing. Of note, Surendranath et al and Decuntom et al concluded that the MinION data quality is improving, but not sufficient yet to be used for routine HLA typing.

Since third generation technologies hold the promise – among others – to resolve phasing ambiguities due to the long reads they generate, we will be interested to hear about further studies that deploy nanopore sequencing in the HLA filed.