P064Evaluation of MIA FORA NGS FLEX HLA typing kit.pdf

P064 EVALUATION OF MIA FORA NGS FLEX HLA TYPING KIT

Ping Rao, Angelica DeOliveira, Candace M. Young, Katherine Leagans, Wendy E. Hanshew, Dong-Feng Chen.

Duke University, Durham, NC, United States

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Aim: As the HLA database expanding, traditional SBT method results in more and more ambiguities and

requires increasing additional testing. We evaluated Mia Fora FLEX11 NGS typing kit with automation intend-

ing to replace SBT with NGS.

Methods: 108 DNA samples were selected. 92 and 16 samples were previously typed by SBT and rSSO/SSP,

respectively. All DNA were prepared from blood except 4 from buccal swab and 3 from B cell line. DNA were

resolution of third field according to WHO nomenclature. 11 ambiguities were identified which were all with

DPB1. 7 potential new alleles were identified: 2 HLA-B, 1 DRB5, 1 DQA1 and 3 DPA1. NGS typing was 99.5%

(1499/1506) concordant with previous typing at allele level. Discordance was observed with DQB1, DRB3/4/5

and DPA1 (Table 1). Allele dropout was observed in 1 out of 23 DQB1*06:02 and 2 out of 11 DRB4*01:01 in the

validation cohort. Both dropouts can be predicted by allele association with DQA1 or DRB1. Sample repeat

rate was 11% (18/161). Four samples failed due to software glitch with the Window 10 version but corrected

when switched to the Linux Version. Most other failure was due to insufficient data which can be resolved by

repeating test. The only failure that could not be resolved by repeat was DRB4*01:01 dropout.

Conclusions: Comparing to SBT method, Mia Fora typing produced high resolution results with ambigui-

ties only presented with DPB1 locus. Little or no additional testing was required. Mia Fora typing had a high