PMID- 28267558
OWN - NLM
STAT- MEDLINE
DCOM- 20170421
LR  - 20220409
IS  - 1879-1166 (Electronic)
IS  - 0198-8859 (Print)
IS  - 0198-8859 (Linking)
VI  - 78
IP  - 4
DP  - 2017 Apr
TI  - Rapid detection of donor cell free DNA in lung transplant recipients with 
      rejections using donor-recipient HLA mismatch.
PG  - 342-349
LID - S0198-8859(17)30039-3 [pii]
LID - 10.1016/j.humimm.2017.03.002 [doi]
AB  - Fiberoptic bronchoscopy and transbronchial lung biopsy are currently the gold 
      standard for detection of acute rejection following human lung transplantation 
      (LTx). However, these surveillance procedures are expensive and invasive. Up to 
      now, there are few new methods that have demonstrated clinical utility for 
      detecting early stages of rejection following human lung transplantation. We 
      optimized and technically validated a novel method to quantify donor-derived 
      circulating cell free DNA (DcfDNA) that can be used as an early biomarker for 
      lung allograft rejection. The method involves the initial development of a panel 
      of probes in which each probe will specifically target a unique sequence of a 
      human leukocyte antigen (HLA) allele. After transplantation, donor/recipient 
      specific probes are chosen based on the mismatched HLA loci, followed by droplet 
      digital PCR (ddPCR) used as a quantitative assay to accurately track the trace 
      amount of DcfDNA in an ample excess of recipient DNA background. The average 
      false positive rate noted was about 1 per 800,000 molecules. Serially 2-fold 
      diluted cfDNA, representing donor fractions of cfDNA, were spiked into a constant 
      level of cfDNA representing the recipient cfDNA. The fraction of spiked cfDNA was 
      measured and quantitative linearity was observed across seven serially diluted 
      cfDNA samples. We were able to measure the minor portion of cfDNA as low as 0.2% 
      of total cfDNA. We subsequently applied the method to a pilot set of 18 LTx 
      recipients grouped into biopsy-proven acute rejection, bronchiolitis obliterans 
      syndrome (BOS) or stable groups. Serial plasma samples were used to identify the 
      percentage of DcfDNA over total cfDNA. The level of DcfDNA was significantly 
      elevated in patients diagnosed with acute rejection (10.30+/-2.80, n=18), compared 
      to that from stable (1.71+/-0.50, n=24) or from BOS patients (2.52+/-0.62, n=20). In 
      conclusion, we present results validating the application of digital PCR to 
      quantify DcfDNA assay in primary clinical specimens, which demonstrate that 
      DcfDNA can be used as an early non-invasive biomarker for acute lung allograft 
      rejection.
CI  - Copyright (c) 2017 American Society for Histocompatibility and Immunogenetics. 
      Published by Elsevier Inc. All rights reserved.
FAU - Zou, Jun
AU  - Zou J
AD  - Department of Laboratory Medicine, University of Texas MD Anderson Cancer Center, 
      Houston, TX 77030, USA. Electronic address: jzou@mdanderson.org.
FAU - Duffy, Brian
AU  - Duffy B
AD  - HLA Laboratory, Barnes-Jewish Hospital, St. Louis, MO 63110, USA.
FAU - Slade, Michael
AU  - Slade M
AD  - Department of Medicine, Washington University School of Medicine, St. Louis, MO 
      63110, USA.
FAU - Young, Andrew Lee
AU  - Young AL
AD  - Department of Pediatrics, Division of Hematology and Oncology, Washington 
      University School of Medicine, Saint Louis, MO, USA; Center for Genome Sciences 
      and Systems Biology, Washington University School of Medicine, Saint Louis, MO, 
      USA.
FAU - Steward, Nancy
AU  - Steward N
AD  - Department of Surgery, Washington University School of Medicine, St. Louis, MO 
      63110, USA.
FAU - Hachem, Ramsey
AU  - Hachem R
AD  - Department of Medicine, Washington University School of Medicine, St. Louis, MO 
      63110, USA.
FAU - Mohanakumar, T
AU  - Mohanakumar T
AD  - Norton Thoracic Institute Research Laboratory, St. Joseph's Hospital & Medical 
      Center, Phoenix, AZ 85013, USA.
LA  - eng
GR  - R01 HL056643/HL/NHLBI NIH HHS/United States
PT  - Journal Article
DEP - 20170304
PL  - United States
TA  - Hum Immunol
JT  - Human immunology
JID - 8010936
RN  - 0 (Biomarkers)
RN  - 0 (HLA Antigens)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Acute Disease
MH  - Adult
MH  - Aged
MH  - Biomarkers/*analysis
MH  - Blood Chemical Analysis/*methods
MH  - Bronchiolitis Obliterans/*diagnosis
MH  - DNA/*analysis
MH  - Diagnosis, Differential
MH  - Female
MH  - Graft Rejection/*diagnosis/genetics
MH  - HLA Antigens/*genetics
MH  - Histocompatibility
MH  - Histocompatibility Testing
MH  - Humans
MH  - *Lung Transplantation
MH  - Male
MH  - Middle Aged
MH  - Tissue Donors
MH  - Transplant Recipients
MH  - Young Adult
PMC - PMC5595356
MID - NIHMS859440
OTO - NOTNLM
OT  - Acute rejection
OT  - Donor cell free DNA
OT  - Droplet digital PCR
OT  - HLA
OT  - Lung transplantation
COIS- All authors have no financial disclosure related to the study described in the 
      submitted manuscript. The authors declare no conflict of interest.
EDAT- 2017/03/08 06:00
MHDA- 2017/04/22 06:00
PMCR- 2018/04/01
CRDT- 2017/03/08 06:00
PHST- 2016/12/16 00:00 [received]
PHST- 2017/02/21 00:00 [revised]
PHST- 2017/03/02 00:00 [accepted]
PHST- 2017/03/08 06:00 [pubmed]
PHST- 2017/04/22 06:00 [medline]
PHST- 2017/03/08 06:00 [entrez]
PHST- 2018/04/01 00:00 [pmc-release]
AID - S0198-8859(17)30039-3 [pii]
AID - 10.1016/j.humimm.2017.03.002 [doi]
PST - ppublish
SO  - Hum Immunol. 2017 Apr;78(4):342-349. doi: 10.1016/j.humimm.2017.03.002. Epub 2017 
      Mar 4.