This study presents an extensive quality assurance (QA) analysis of Next-Generation Sequencing (NGS) in a clinical HLA laboratory.

Samples were sequenced for 11 HLA genes by Omixon Holotype NGS Assay and confirmatory testing for QA purposes was performed using OneLambda LABTypeRSSO or MicroSSP to confirm accuracy. 

1054 sequenced samples were analyzed and confirmatory testing was performed in 8.3% of them. Of the 87 analytical query tests, 51.7% arose from a potential drop out, and 21.7% verified a true drop out: an allele not detected by NGS. 25.8% of analytical queries were due to questionable QA metrics. Compared to the previous version of the Omixon kit (n=1180), dropout testing decreased to 4.2%, QC metric testing to 2.0%, and the detection of true dropouts to 0.9%. 

Advancements in NGS chemistry have enabled an important decrease in confirmatory typing and support NGS as a reliable technology to produce high-resolution genotypes needed for epitope-matching.

QUALITY ANALYSIS OF NGS IN GENERATING HLA GENOTYPES IN A CLINICAL LAB