PMID- 31669229
OWN - NLM
STAT- MEDLINE
DCOM- 20210510
LR  - 20221207
IS  - 1943-7811 (Electronic)
IS  - 1525-1578 (Linking)
VI  - 22
IP  - 1
DP  - 2020 Jan
TI  - Using Nanopore Whole-Transcriptome Sequencing for Human Leukocyte Antigen 
      Genotyping and Correlating Donor Human Leukocyte Antigen Expression with Flow 
      Cytometric Crossmatch Results.
PG  - 101-110
LID - S1525-1578(19)30404-0 [pii]
LID - 10.1016/j.jmoldx.2019.09.005 [doi]
AB  - Transplant centers are increasingly using virtual crossmatching to evaluate 
      recipient and donor compatibility. However, the current state of virtual 
      crossmatching fails to incorporate donor human leukocyte antigen (HLA) expression 
      in the assessment, despite numerous studies that have demonstrated the impact of 
      donor HLA expression on physical crossmatch outcomes. Whole-transcriptome 
      sequencing (RNA-Seq) for HLA enables simultaneous determination of HLA genotyping 
      and relative HLA expression. Ultimately the RNA-Seq needs to be faster to be 
      incorporated into the virtual crossmatching process. However, to demonstrate 
      feasibility, the utility of the MinION sequencer (Oxford Nanopore Technologies, 
      Oxford, UK) was evaluated in combination with RNA-Seq to generate HLA genotypes 
      and to determine HLA class I expression. Although HLA class I expression varied 
      among individuals, the pattern of HLA expression remained relatively consistent 
      (HLA-B > HLA-A = HLA-C). HLA-A and -C had similar expression profiles. The impact 
      of donor HLA expression was evaluated using serum samples containing a single 
      donor-specific antibody (DSA). By making DSA consistent, donor HLA expression 
      variability could be assessed. With consistent DSA mean fluorescence intensity, 
      there was a direct relationship between the donor HLA expression to which the DSA 
      is against and flow cytometric crossmatch median channel shifts.
CI  - Copyright (c) 2020 American Society for Investigative Pathology and the Association 
      for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
FAU - Montgomery, Maureen C
AU  - Montgomery MC
AD  - Molecular Immunology Laboratory, McLendon Clinical Laboratories, University of 
      North Carolina Hospitals, Chapel Hill, North Carolina.
FAU - Liu, Chang
AU  - Liu C
AD  - Department of Pathology and Immunology, Washington University School of Medicine, 
      St. Louis, Missouri.
FAU - Petraroia, Rosanne
AU  - Petraroia R
AD  - HLA Laboratory, McLendon Clinical Laboratories, University of North Carolina 
      Hospitals, Chapel Hill, North Carolina.
FAU - Weimer, Eric T
AU  - Weimer ET
AD  - Molecular Immunology Laboratory, McLendon Clinical Laboratories, University of 
      North Carolina Hospitals, Chapel Hill, North Carolina; HLA Laboratory, McLendon 
      Clinical Laboratories, University of North Carolina Hospitals, Chapel Hill, North 
      Carolina; Department of Pathology & Laboratory Medicine, University of North 
      Carolina at Chapel Hill School of Medicine, Chapel Hill, North Carolina. 
      Electronic address: eric_weimer@med.unc.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20191024
PL  - United States
TA  - J Mol Diagn
JT  - The Journal of molecular diagnostics : JMD
JID - 100893612
RN  - 0 (Antibodies)
RN  - 0 (HLA Antigens)
RN  - 0 (Histocompatibility Antigens Class I)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Antibodies/immunology
MH  - Blood Donors
MH  - Cells, Cultured
MH  - Flow Cytometry/*methods
MH  - *Genotype
MH  - Genotyping Techniques
MH  - HLA Antigens/*genetics/immunology
MH  - Histocompatibility Antigens Class I/*genetics/immunology
MH  - Histocompatibility Testing/*methods
MH  - Humans
MH  - Lymphocytes/metabolism
MH  - *Nanopores
MH  - Organ Transplantation
MH  - RNA, Messenger/genetics/isolation & purification
MH  - RNA-Seq
MH  - *Tissue Donors
MH  - Transplant Recipients
MH  - Exome Sequencing/*methods
EDAT- 2019/11/02 06:00
MHDA- 2021/05/11 06:00
CRDT- 2019/11/01 06:00
PHST- 2019/07/26 00:00 [received]
PHST- 2019/08/27 00:00 [revised]
PHST- 2019/09/11 00:00 [accepted]
PHST- 2019/11/02 06:00 [pubmed]
PHST- 2021/05/11 06:00 [medline]
PHST- 2019/11/01 06:00 [entrez]
AID - S1525-1578(19)30404-0 [pii]
AID - 10.1016/j.jmoldx.2019.09.005 [doi]
PST - ppublish
SO  - J Mol Diagn. 2020 Jan;22(1):101-110. doi: 10.1016/j.jmoldx.2019.09.005. Epub 2019 
      Oct 24.