IL8 and EDEM3 gene expression ratio indicates peripheral blood mononuclear cell (PBMC) quality.
PBMC
Preanalytical
Quality control
Journal
Journal of immunological methods
ISSN: 1872-7905
Titre abrégé: J Immunol Methods
Pays: Netherlands
ID NLM: 1305440
Informations de publication
Date de publication:
02 2019
02 2019
Historique:
received:
27
03
2018
revised:
19
11
2018
accepted:
19
11
2018
pubmed:
30
11
2018
medline:
16
1
2020
entrez:
30
11
2018
Statut:
ppublish
Résumé
Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. We used Taqman RTqPCR to evaluate the combination of two target genes for their "diagnostic performance" in identifying EDTA and citrate-anticoagulated PBMC samples with extended pre-centrifugation times. We established the PBMC preanalytical score, a gene expression metric to asses the PBMC quality related to the pre-centrifugation delay at room temperature for different anticoagulants. The PBMC preanalytical score measurement can identify: CONCLUSION: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.
Sections du résumé
BACKGROUND
Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs).
METHODS
PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. We used Taqman RTqPCR to evaluate the combination of two target genes for their "diagnostic performance" in identifying EDTA and citrate-anticoagulated PBMC samples with extended pre-centrifugation times.
RESULTS
We established the PBMC preanalytical score, a gene expression metric to asses the PBMC quality related to the pre-centrifugation delay at room temperature for different anticoagulants. The PBMC preanalytical score measurement can identify: CONCLUSION: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.
Identifiants
pubmed: 30496732
pii: S0022-1759(18)30118-2
doi: 10.1016/j.jim.2018.11.012
pii:
doi:
Substances chimiques
CXCL8 protein, human
0
Calcium-Binding Proteins
0
Interleukin-8
0
EDEM3 protein, human
EC 3.2.1.24
alpha-Mannosidase
EC 3.2.1.24
Types de publication
Clinical Trial
Journal Article
Multicenter Study
Langues
eng
Sous-ensembles de citation
IM
Pagination
13-19Commentaires et corrections
Type : ErratumIn
Informations de copyright
Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.