Single-Tube Polymerase Chain Reaction Amplification Protocol for Sequencing-Based Typing of Human Leukocyte Antigen DRB1 / 中国普外基础与临床杂志

ABSTRACT

Objective To develop a new single-tube polymerase chain reaction amplification (ST Amp) protocol for the efficient sequencing-based typing (SBT) of human leukocyte antigen DRB1(HLA-DRB1).Methods A set of 7 group-specific exonic 5′ amplification primers and a single generic 3′ primer were included together in a single PCR mix to facilitate a single PCR amplification per sample for HLA-DRB1 typing.Results All samples were successfully typed, the typing result was accurate and repeatable.Conclusion ST Amp technique has resulted in the ability to perform high-resolution, high-specificity and high-throughput HLA-DRB1 typing by DNA sequencing.