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Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing / 대한수혈학회지
Korean Journal of Blood Transfusion ; : 310-319, 2018.
Article in Korean | WPRIM | ID: wpr-718424
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.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Volunteers / Bone Marrow / Histocompatibility Testing / HLA-A Antigens / Gene Library / Polymerase Chain Reaction / Statistics as Topic / Computational Biology / Methods Language: Korean Journal: Korean Journal of Blood Transfusion Year: 2018 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Volunteers / Bone Marrow / Histocompatibility Testing / HLA-A Antigens / Gene Library / Polymerase Chain Reaction / Statistics as Topic / Computational Biology / Methods Language: Korean Journal: Korean Journal of Blood Transfusion Year: 2018 Type: Article