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Use of quantitative real time polymerase chain reaction to assess gene transcripts associated with antibody-mediated rejection of kidney transplants

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Title: Use of quantitative real time polymerase chain reaction to assess gene transcripts associated with antibody-mediated rejection of kidney transplants
Authors: Dominy, KM
Roufosse, C
De Kort, H
Willicombe, M
Brookes, P
Behmoaras, JV
Petretto, EG
Galliford, J
Choi, P
Taube, D
Cook, HT
Mclean, AG
Item Type: Journal Article
Abstract: Introduction Microarray studies have shown elevated transcript levels of endothelial and natural killer (NK) cell–associated genes during antibody-mediated rejection (AMR) of the renal allograft. This study aimed to assess the use of quantitative real-time polymerase chain reaction as an alternative to microarray analysis on a subset of these elevated genes. Methods Thirty-nine renal transplant biopsies from patients with de novo donor-specific antibodies and eighteen 1-year surveillance biopsies with no histological evidence of rejection were analyzed for expression of 11 genes previously identified as elevated in AMR. Results Expression levels of natural killer markers were correlated to microcirculation inflammation and graft outcomes to a greater extent than endothelial markers. Creating a predictive model reduced the number of gene transcripts to be assessed to 2, SH2D1b and MYBL1, resulting in 66.7% sensitivity and 89.7% specificity for graft loss. Discussion This work demonstrates that elevated gene expression levels, proposed to be associated with AMR, can be detected by established quantitative real-time polymerase chain reaction technology, making transition to the clinical setting feasible. Transcript analysis provides additional diagnostic information to the classification schema for AMR diagnosis but it remains to be determined whether significant numbers of centres will validate transcript analysis in their laboratories and put such analysis into clinical use.
Issue Date: 1-Sep-2015
Date of Acceptance: 19-Nov-2014
URI: http://hdl.handle.net/10044/1/53841
DOI: https://dx.doi.org/10.1097/TP.0000000000000621
ISSN: 0041-1337
Publisher: Lippincott, Williams & Wilkins
Start Page: 1981
End Page: 1988
Volume: 99
Issue: 9
Copyright Statement: © 2015 Wolters Kluwer Health, Inc. All rights reserved.
Sponsor/Funder: Wellcome Trust
Medical Research Council (MRC)
Funder's Grant Number: 092523/Z/10/Z
MR/M004716/1
Keywords: Science & Technology
Life Sciences & Biomedicine
Immunology
Surgery
Transplantation
DONOR-SPECIFIC ANTIBODIES
RENAL-ALLOGRAFT PATHOLOGY
MICROCIRCULATION INFLAMMATION
SUBCLINICAL REJECTION
BIOPSIES
PCR
CLASSIFICATION
EXPRESSION
RECIPIENTS
DIAGNOSIS
Biopsy
Case-Control Studies
Gene Expression Profiling
Genetic Markers
Graft Rejection
Graft Survival
Humans
Immunity, Humoral
Kaplan-Meier Estimate
Kidney Transplantation
Predictive Value of Tests
Proto-Oncogene Proteins
RNA, Messenger
Real-Time Polymerase Chain Reaction
Risk Factors
Trans-Activators
Transcription Factors
Transcription, Genetic
Treatment Outcome
11 Medical And Health Sciences
Publication Status: Published
Appears in Collections:Clinical Sciences
Molecular Sciences
Department of Medicine



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