PMID- 27543980
OWN - NLM
STAT- MEDLINE
DCOM- 20170306
LR  - 20220311
IS  - 1943-7722 (Electronic)
IS  - 0002-9173 (Linking)
VI  - 146
IP  - 3
DP  - 2016 Sep
TI  - EDTA Treatment of Serum Unmasks Complement-Mediated Prozone Inhibition in Human 
      Leukocyte Antigen Antibody Testing.
PG  - 346-52
LID - 10.1093/ajcp/aqw116 [doi]
AB  - OBJECTIVES: Luminex-based single-antigen bead human leukocyte antigen (HLA) 
      antibody testing is widely used to define HLA antibodies for transplant 
      compatibility. False-negative results can occur with complement-mediated prozone 
      inhibition. This study assessed the effect of EDTA on the assay background 
      reactivity and fluctuations in antibody mean fluorescent intensity. METHODS: 
      Serum specimens were retrospectively tested using Luminex-based single-antigen 
      beads with and without EDTA. Treated and untreated serum samples were compared by 
      two measures: changes in background reactivity and changes in HLA antibody 
      strength after EDTA treatment. RESULTS: Ten pretransplant and 48 posttransplant 
      specimens were identified: lung (22), heart (10), kidney (21), heart/lung (two), 
      pancreas (one), small bowel (one), and liver (one). After EDTA treatment, weak 
      antibodies (below 2,000 mean florescent intensity) demonstrated the largest 
      fluctuations. Newly identified HLA antibodies were seen in 16% (8/49) of class I 
      and 26% (15/57) of class II beads. EDTA treatment did not result in 
      false-negative reactions compared with untreated serum. CONCLUSIONS: EDTA serum 
      pretreatment mitigated complement-mediated prozone inhibition and improved 
      accurate HLA antibody detection. The background reactivity and the false-negative 
      rate of the assay appear unchanged.
CI  - (c) American Society for Clinical Pathology, 2016. All rights reserved. For 
      permissions, please e-mail: journals.permissions@oup.com.
FAU - Anani, Waseem Q
AU  - Anani WQ
AD  - From the Department of Pathology, University of Pittsburgh Medical Center, 
      Pittsburgh, PA.
FAU - Zeevi, Adriana
AU  - Zeevi A
AD  - From the Department of Pathology, University of Pittsburgh Medical Center, 
      Pittsburgh, PA.
FAU - Lunz, John G
AU  - Lunz JG
AD  - From the Department of Pathology, University of Pittsburgh Medical Center, 
      Pittsburgh, PA. jlunz@giftofhope.org.
LA  - eng
PT  - Journal Article
PL  - England
TA  - Am J Clin Pathol
JT  - American journal of clinical pathology
JID - 0370470
RN  - 0 (HLA Antigens)
RN  - 9007-36-7 (Complement System Proteins)
RN  - 9G34HU7RV0 (Edetic Acid)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Aged
MH  - Child
MH  - Child, Preschool
MH  - Complement System Proteins/*immunology
MH  - Edetic Acid
MH  - Graft Rejection/immunology
MH  - HLA Antigens/*immunology
MH  - Histocompatibility Testing/*methods
MH  - Humans
MH  - Infant
MH  - Middle Aged
MH  - Organ Transplantation/*methods
MH  - Retrospective Studies
MH  - Young Adult
OTO - NOTNLM
OT  - C1q
OT  - EDTA
OT  - complement
OT  - luminex single antigen bead
OT  - prozone effect
OT  - prozone inhibition
EDAT- 2016/08/21 06:00
MHDA- 2017/03/07 06:00
CRDT- 2016/08/21 06:00
PHST- 2016/08/21 06:00 [entrez]
PHST- 2016/08/21 06:00 [pubmed]
PHST- 2017/03/07 06:00 [medline]
AID - aqw116 [pii]
AID - 10.1093/ajcp/aqw116 [doi]
PST - ppublish
SO  - Am J Clin Pathol. 2016 Sep;146(3):346-52. doi: 10.1093/ajcp/aqw116.