Korean J Blood Transfus.
2018 Dec;29(3):310-319. 10.17945/kjbt.2018.29.3.310.
Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing
- Affiliations
-
- 1LabGenomics Clinical Laboratories, Seongnam, Korea. seo2023@nate.com
- KMID: 2435625
- DOI: http://doi.org/10.17945/kjbt.2018.29.3.310
Abstract
- BACKGROUND
Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time.
METHODS
For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing.
RESULTS
DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples.
CONCLUSION
The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.
Keyword
- HLA; NGS; HLA typing
