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KMID : 0387720180290030310
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Korean Journal of Blood Transfusion
2018 Volume.29 No. 3 p.310 ~ p.319
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Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing
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Seo Dong-Hee
Lee Jeong-Min
Park Mi-Ok
Lee Hyun-Ju
Moon Seo-Yoon
Oh Mi-Jin
Kim So-Young
Lee Sang-Heon
Hyeong Ki-Eun
Hu Hae-Jin
Cho Dae-Yeon
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Abstract
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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.
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KEYWORD
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HLA, NGS, HLA typing
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