Recently, HLA epitopes on donor HLA molecules have been shown to be important in the success of solid organ transplantation. However, these epitopes can only be defined using high‐resolution typing results of which are often not available prior to deceased donor allocation. The ability to perform high‐resolution typing at all HLA loci for deceased organ donor allocation prior to transplantation would have major clinical benefits, in particular for highly sensitised recipients. We, therefore, developed a rapid high‐resolution next generation sequencing (NGS) HLA typing (ONT‐Rapid HR HLA) method for on‐call deceased donor allocation using the AllType 11 loci single‐tube assay (OneLambda Inc), modified in‐house to reduce polymerase chain reaction amplification time, and the Oxford Nanopore single‐molecule sequencing platform on the Flongle flow cell. The ONT‐Rapid HR HLA method was validated on 42 samples previously typed by current on‐call sequence‐specific oligonucleotide (HistoSpot) and NGS methods (AllType/Ion Torrent). High‐resolution typing obtained using the ONT‐Rapid HR HLA typing method was 100% concordant with both the current SSO and NGS methods, and in some cases, obtained higher resolution than either of the current methods. The rapid ONT‐Rapid HR HLA typing method was able to obtain these typing results at all loci in 4 to 4.5 hours. The novel ONT‐Rapid HR HLA typing method is the first reported NGS HLA typing method utilised for deceased donor allocation. The ability to provide high‐resolution HLA typing on deceased donors before implantation will in the future allow improvements in matching, which will ultimately provide clinical benefits to patients.

中文翻译：

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利用MinION Oxford纳米孔测序技术进行的高分辨率高分辨率HLA基因分型，用于已故供体器官的分配。

最近，已证明供体HLA分子上的HLA表位对于固体器官移植的成功很重要。但是，只能使用高分辨率的分型结果来定义这些表位，而在捐赠者去世之前，通常无法获得其高分辨率分型结果。能够在所有HLA基因座上进行高分辨率分型，以便在移植前分配已故的器官供体，这将具有重大的临床益处，特别是对于高度敏感的接受者。因此，我们使用AllType 11基因座单管测定法（OneLambda Inc）开发了一种快速高分辨率的下一代测序（NGS）HLA分型（ONT-Rapid HR HLA）方法，用于待命死亡的供体分配，并在可以缩短聚合酶链反应的扩增时间，并在Flongle流通池上使用牛津纳米孔单分子测序平台。在先前使用当前呼叫序列特异性寡核苷酸（HistoSpot）和NGS方法（AllType / Ion Torrent）进行分型的42个样品上，对ONT-Rapid HR HLA方法进行了验证。使用ONT-Rapid HR HLA分型方法获得的高分辨率分型与当前的SSO和NGS方法均100％一致，并且在某些情况下，比任何一种当前方法都可获得更高的分辨率。快速的ONT‐Rapid HR HLA分型方法能够在4到4.5小时内在所有位点获得这些分型结果。新颖的ONT快速HR HLA分型方法是第一个报告的NGS HLA分型方法，用于已故的捐赠者分配。将来在植入前在已故的供体上提供高分辨率HLA分型的能力将使匹配度得到改善，最终将为患者带来临床益处。