Comparison of multiplex cytokine assays in a pediatric cohort with epilepsy.
Chemokines
Cytokines
Luminex
Meso-scale discovery
Neuro-inflammation
Pediatric epilepsy
Journal
Heliyon
ISSN: 2405-8440
Titre abrégé: Heliyon
Pays: England
ID NLM: 101672560
Informations de publication
Date de publication:
Mar 2021
Mar 2021
Historique:
received:
29
08
2020
revised:
12
01
2021
accepted:
04
03
2021
entrez:
22
3
2021
pubmed:
23
3
2021
medline:
23
3
2021
Statut:
epublish
Résumé
Multiplex analyses allow for detection of dozens of cytokines/chemokines in small sample volumes. Although several commercially available assay kits are available, there are no comparative data in plasma measurements among pediatric or epilepsy cohorts. Cohort study of 38 children with epilepsy. We evaluated plasma levels of cytokines/chemokines using three different assays: Luminex® xMAP high-sensitivity (HS) and standard-sensitivity (SS) assays, and Meso-Scale Discovery (MSD). We calculated recovery rates of each analyte, correlation coefficients between assays, and level of agreement between measurements. We repeated analyses in a subset of samples after a single freeze-thaw cycle. Among ten analytes common to all assays, HS had high recovery (<15% of values extrapolated or out-of- range [OOR]) for all analytes, SS for 50%, and MSD for 40%. While several analytes had a high correlation between assays, Bland-Altman plots demonstrated assays were not interchangeable. For most analytes, a single freeze-thaw cycle decreased cytokines/chemokine measurements. There was good correlation of measurements after a freeze-thaw cycle with acceptable agreement between measurements for six of 13 (46%) analytes using HS, one of 9 (11%) for SS, and none for MSD. HS assays may optimize yield in plasma for proteins of particular interest in epilepsy research, limit values extrapolated beyond the standard curve, and improve precision compared to other SS and MSD assays. Our results demonstrate assay choice may be critical to study results and support the need for a standardized approach to biomarker assessment across epilepsy research and other domains.
Sections du résumé
BACKGROUND
BACKGROUND
Multiplex analyses allow for detection of dozens of cytokines/chemokines in small sample volumes. Although several commercially available assay kits are available, there are no comparative data in plasma measurements among pediatric or epilepsy cohorts.
NEW METHOD
METHODS
Cohort study of 38 children with epilepsy. We evaluated plasma levels of cytokines/chemokines using three different assays: Luminex® xMAP high-sensitivity (HS) and standard-sensitivity (SS) assays, and Meso-Scale Discovery (MSD). We calculated recovery rates of each analyte, correlation coefficients between assays, and level of agreement between measurements. We repeated analyses in a subset of samples after a single freeze-thaw cycle.
RESULTS
RESULTS
Among ten analytes common to all assays, HS had high recovery (<15% of values extrapolated or out-of- range [OOR]) for all analytes, SS for 50%, and MSD for 40%. While several analytes had a high correlation between assays, Bland-Altman plots demonstrated assays were not interchangeable. For most analytes, a single freeze-thaw cycle decreased cytokines/chemokine measurements. There was good correlation of measurements after a freeze-thaw cycle with acceptable agreement between measurements for six of 13 (46%) analytes using HS, one of 9 (11%) for SS, and none for MSD.
COMPARISON WITH EXISTING METHODS
METHODS
HS assays may optimize yield in plasma for proteins of particular interest in epilepsy research, limit values extrapolated beyond the standard curve, and improve precision compared to other SS and MSD assays.
CONCLUSION
CONCLUSIONS
Our results demonstrate assay choice may be critical to study results and support the need for a standardized approach to biomarker assessment across epilepsy research and other domains.
Identifiants
pubmed: 33748497
doi: 10.1016/j.heliyon.2021.e06445
pii: S2405-8440(21)00550-8
pmc: PMC7966851
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e06445Subventions
Organisme : NINDS NIH HHS
ID : K23 NS105918
Pays : United States
Informations de copyright
© 2021 The Authors.
Déclaration de conflit d'intérêts
The authors declare the following conflict of interests: Dr. Numis reports Grant Support from the 10.13039/100000002NIH (K23NS105918). Dr. Fox reports Grant Support from the 10.13039/100000002NIH (U54NS065705) and the Pediatric Epilepsy Research Foundation.
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