Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing.
10.17945/kjbt.2018.29.3.310
- Author:
Dong Hee SEO
1
;
Jeong Min LEE
;
Mi Ok PARK
;
Hyun Ju LEE
;
Seo Yoon MOON
;
Mijin OH
;
So Young KIM
;
Sang Heon LEE
;
Ki Eun HYEONG
;
Hae Jin HU
;
Dae Yeon CHO
Author Information
1. LabGenomics Clinical Laboratories, Seongnam, Korea. seo2023@nate.com
- Publication Type:Original Article
- Keywords:
HLA;
NGS;
HLA typing
- MeSH:
Bone Marrow;
Computational Biology;
Gene Library;
Histocompatibility Testing;
HLA-A Antigens*;
Methods*;
Polymerase Chain Reaction;
Statistics as Topic;
Volunteers
- From:Korean Journal of Blood Transfusion
2018;29(3):310-319
- CountryRepublic of Korea
- Language:Korean
-
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.
