Long-Read Next-Generation Sequencing Technologies Can Address Some Limitations of Short-Read Technologies in HLA...

Area of Science:

• Immunogenetics
• Genomic Sequencing Technologies

Background:

• Accurate Human Leukocyte Antigen (HLA) allele identification is critical for successful graft-recipient matching in transplantation.
• Next-generation sequencing (NGS) has advanced HLA typing resolution, but challenges remain in precise allele assignment.

Purpose of the Study:

• To investigate a novel Human Leukocyte Antigen (HLA)-C*12 allele identified in a bone marrow donor.
• To evaluate the utility of long-read sequencing in resolving complex HLA typing ambiguities.

Main Methods:

• Utilized short-read and long-read next-generation sequencing (NGS) for Human Leukocyte Antigen (HLA) typing.
• Performed Sanger sequencing on parental samples to confirm the origin of a novel mutation.

Main Results:

• A novel Human Leukocyte Antigen (HLA)-C*12 allele with a silent G-to-A mutation was identified.
• Short-read NGS initially linked the mutation to HLA-C*12:03:01, but long-read sequencing accurately assigned it to HLA-C*12:02:02.
• Long-read sequencing successfully resolved cis-trans ambiguities and improved allele phasing.

Conclusions:

• Long-read sequencing offers superior accuracy for new Human Leukocyte Antigen (HLA) allele identification.
• This technology enhances the ability to resolve complex genetic phasing and allelic ambiguities in HLA typing.
• Improved HLA typing accuracy is essential for ensuring graft-recipient compatibility.