P179 Multi-center study on automating holotype HLA on a Biomek 4000
2017; Elsevier BV; Volume: 78; Linguagem: Inglês
10.1016/j.humimm.2017.06.239
ISSN1879-1166
AutoresEfi Melista, Kim McGowan, Doreen Sese, Sharon Adams, Deborah Chen, Tünde Vágó, Zach Smith, Shilpa Parakh, Peter Meintjes,
Tópico(s)Renal Transplantation Outcomes and Treatments
ResumoHolotype HLA is a powerful NGS-based method for HLA genotyping being adopted by HLA laboratories over legacy technologies such as SSO and Sanger SBT for its increased accuracy, resolution, repeatability and high-throughput nature. The elimination of human and random error is a desirable attribute of any diagnostic test and the implementation of a liquid handling system can significantly reduce both sources of error in an NGS workflow. Development of a protocol that streamlines multiple modules of the NGS process was the goal of this multi-center study. The Beckman Coulter Biomek 4000 automated liquid handler is a small footprint solution for pre- and post-PCR processes in NGS-based HLA typing. During development, the protocol was optimized through comparison of multiple data points at measurable stopping points to provide a customized modular liquid handling solution for any HLA lab. Technologists and scientists from three sites performed multiple runs of 24 samples for 11 HLA loci using the Holotype HLA 24/11 configuration automated on the Biomek 4000 to generate data on the accuracy, specificity, repeatability and reproducibility of an automated Holotype HLA solution. Data obtained from the three centers were analyzed using Omixon HLA Twin and assessed for accuracy, reproducibility, and allele assignment ambiguities to demonstrate that high-throughput NGS-based HLA typing is achievable in under 48 h from gDNA to genotype using the Beckman Coulter Biomek 4000 modular based NGS protocol for Holotype HLA 24/11.
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