Qu Zhengzhong James
Genome Institute of Singapore, Singapore
Title: Development of a comprehensive high-coverage, cost effective HLA typing assay
Biography
Biography: Qu Zhengzhong James
Abstract
The human leukocyte antigen (HLA) is responsible for regulation of the immune system in humans. HLA typing has important clinical significance for tissue transplantation matching, autoimmune disease-association studies and drug hypersensitivity research. However, the highly polymorphic essence of the HLA genes makes it a challenging task. Conventional methods like SSO and SSP only provide lower resolution typing results and cannot identify new alleles. High-resolution techniques like SBT required other methods for confirmation of phasing and it is time consuming. Single Molecule Real Time (SMRT) sequencing is a parallelized single molecule DNA sequencing by synthesis technology developed by Pacific Biosciences. It has long read lengths which enable unambiguous phasing of the entire HLA genes. However, the cost of the SMRT sequencing is still too high for clinical routine use. To develop clinically implementable HLA typing assay by using SMRT sequencing technology, we designed two sets of multiplex PCR primers for the HLA genes. The first assay we developed enables the HLA typing of A, B, C, DRB1, DQB1, DQA1, DPA1 and DPB1 genes for one patient by using one SMRT cell. The second assay we developed enables the typing of HLA-A genes for up to 50 patients by using one SMRT cell. Our assays provided choice of both coverage and cost-effectiveness.