Two-field resolution on-call HLA typing for deceased donors using nanopore sequencing

Area of Science:

• Immunogenetics
• Transplantation immunology
• Molecular diagnostics

Background:

• Current HLA genotyping methods for deceased donors face time and resolution limitations.
• Accurate HLA compatibility assessment is crucial for anti-HLA sensitized recipients, especially with allele-specific antibodies.
• Timely and precise HLA data is essential for organ allocation decisions.

Purpose of the Study:

• To develop and validate a rapid, high-resolution HLA genotyping protocol for deceased donors.
• To assess the feasibility of using nanopore sequencing for real-time HLA typing in deceased organ donation.
• To improve organ allocation security for highly sensitized recipients.

Main Methods:

• A customized protocol using the NanoTYPE® HLA typing kit and MinION® sequencer was developed.
• Prospective real-time analysis of HLA typing was performed on 18 deceased donors.
• The nanopore-based method's speed and accuracy were compared to conventional approaches.

Main Results:

• The protocol achieved average HLA typing in 3.75 hours with three-field resolution and minimal ambiguity.
• The method's duration was compatible with organ donation and allocation timelines.
• The enhanced resolution improved the security of organ allocation, particularly for complex cases.

Conclusions:

• Nanopore-based HLA typing offers a rapid and accurate solution for deceased donors.
• This approach enhances organ allocation safety and efficiency, especially for vulnerable recipients.
• The protocol has the potential to significantly impact deceased donor HLA genotyping practices.