PMID- 27376474
OWN - NLM
STAT- MEDLINE
DCOM- 20170606
LR  - 20180122
IS  - 1943-7811 (Electronic)
IS  - 1525-1578 (Linking)
VI  - 18
IP  - 5
DP  - 2016 Sep
TI  - Performance Characteristics and Validation of Next-Generation Sequencing for 
      Human Leucocyte Antigen Typing.
PG  - 668-675
LID - S1525-1578(16)30065-4 [pii]
LID - 10.1016/j.jmoldx.2016.03.009 [doi]
AB  - High-resolution human leukocyte antigen (HLA) matching reduces graft-versus-host 
      disease and improves overall patient survival after hematopoietic stem cell 
      transplant. Sanger sequencing has been the gold standard for HLA typing since 
      1996. However, given the increasing number of new HLA alleles identified and the 
      complexity of the HLA genes, clinical HLA typing by Sanger sequencing requires 
      several rounds of additional testing to provide allele-level resolution. Although 
      next-generation sequencing (NGS) is routinely used in molecular genetics, few 
      clinical HLA laboratories use the technology. The performance characteristics of 
      NGS HLA typing using TruSight HLA were determined using Sanger sequencing as the 
      reference method. In total, 211 samples were analyzed with an overall accuracy of 
      99.8% (2954/2961) and 46 samples were analyzed for precision with 100% (368/368) 
      reproducibility. Most discordant alleles were because of technical error rather 
      than assay performance. More important, the ambiguity rate was 3.5% (103/2961). 
      Seventy-four percentage of the ambiguities were within the DRB1 and DRB4 loci. 
      HLA typing by NGS saves approximately $6000 per run when compared to Sanger 
      sequencing. Thus, TruSight HLA assay enables high-throughput HLA typing with an 
      accuracy, precision, ambiguity rate, and cost savings that should facilitate 
      adoption of NGS technology in clinical HLA laboratories.
CI  - Copyright (c) 2016 American Society for Investigative Pathology and the Association 
      for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
FAU - Weimer, Eric T
AU  - Weimer ET
AD  - Human Leukocyte Antigen Laboratory, McLendon Clinical Laboratories, University of 
      North Carolina Hospitals, Chapel Hill, North Carolina; Department of Pathology 
      and Laboratory Medicine, University of North Carolina at Chapel Hill School of 
      Medicine, Chapel Hill, North Carolina. Electronic address: 
      eric_weimer@med.unc.edu.
FAU - Montgomery, Maureen
AU  - Montgomery M
AD  - Human Leukocyte Antigen Laboratory, McLendon Clinical Laboratories, University of 
      North Carolina Hospitals, Chapel Hill, North Carolina.
FAU - Petraroia, Rosanne
AU  - Petraroia R
AD  - Human Leukocyte Antigen Laboratory, McLendon Clinical Laboratories, University of 
      North Carolina Hospitals, Chapel Hill, North Carolina.
FAU - Crawford, John
AU  - Crawford J
AD  - Human Leukocyte Antigen Laboratory, McLendon Clinical Laboratories, University of 
      North Carolina Hospitals, Chapel Hill, North Carolina.
FAU - Schmitz, John L
AU  - Schmitz JL
AD  - Human Leukocyte Antigen Laboratory, McLendon Clinical Laboratories, University of 
      North Carolina Hospitals, Chapel Hill, North Carolina; Department of Pathology 
      and Laboratory Medicine, University of North Carolina at Chapel Hill School of 
      Medicine, Chapel Hill, North Carolina.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160701
PL  - United States
TA  - J Mol Diagn
JT  - The Journal of molecular diagnostics : JMD
JID - 100893612
RN  - 0 (HLA Antigens)
SB  - IM
EIN - J Mol Diagn. 2016 Nov;18(6):933. PMID: 27657454
MH  - Cost-Benefit Analysis
MH  - Gene Amplification
MH  - Gene Library
MH  - HLA Antigens/*genetics
MH  - *High-Throughput Nucleotide Sequencing/economics/methods/standards
MH  - Histocompatibility Testing/economics/*methods/standards
MH  - Humans
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Sequence Analysis, DNA/methods/standards
EDAT- 2016/07/05 06:00
MHDA- 2017/06/07 06:00
CRDT- 2016/07/05 06:00
PHST- 2015/12/17 00:00 [received]
PHST- 2016/02/29 00:00 [revised]
PHST- 2016/03/23 00:00 [accepted]
PHST- 2016/07/05 06:00 [entrez]
PHST- 2016/07/05 06:00 [pubmed]
PHST- 2017/06/07 06:00 [medline]
AID - S1525-1578(16)30065-4 [pii]
AID - 10.1016/j.jmoldx.2016.03.009 [doi]
PST - ppublish
SO  - J Mol Diagn. 2016 Sep;18(5):668-675. doi: 10.1016/j.jmoldx.2016.03.009. Epub 2016 
      Jul 1.