Document Detail


Development and validation of a multiplex microsphere-based assay for detection of domestic cat (Felis catus) cytokines.
MedLine Citation:
PMID:  21209158     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cytokines are essential signaling molecules that mediate the innate immune response, and therefore their presence can be of diagnostic, prognostic, and pathogenic significance. Microsphere-based immunoassays allow rapid and accurate evaluation of cytokine levels in several species, including humans, dogs, and mice; however, technology to evaluate domestic cat (Felis catus) cytokines has been limited to single-analyte enzyme-linked immunosorbent assays (ELISAs). Microsphere-based immunoassays provide an attractive alternative technology for detecting and quantifying multiple analytes in a single assay using as little as 50 μl of sample. We describe the development and validation of a microsphere-based assay for three commonly analyzed domestic cat cytokines (gamma interferon, interleukin-10, and interleukin-12/interleukin-23 p40) using reagents from commercially available ELISAs. The assay was optimized for capture and detection antibody concentrations, streptavidin-phycoerythrin concentration, and number of microspheres. The validated lower and upper quantitation limits were 31 and 1,000 pg/ml for gamma interferon, 63 and 2,000 pg/ml for interleukin-10, and 39 and 625 pg/ml for interleukin-12/interleukin-23 p40. Cytokine concentrations in peripheral blood mononuclear cell supernatants were measured, and results obtained by the microsphere assay were correlated with values obtained with commercially available ELISA kits. This technology is a convenient and reproducible assay to evaluate domestic cat cytokine responses elicited by a variety of diseases.
Authors:
Britta A Wood; Kevin P O'Halloran; Sue Vandewoude
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural     Date:  2011-01-05
Journal Detail:
Title:  Clinical and vaccine immunology : CVI     Volume:  18     ISSN:  1556-679X     ISO Abbreviation:  Clin. Vaccine Immunol.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-03-03     Completed Date:  2011-06-07     Revised Date:  2011-09-13    
Medline Journal Info:
Nlm Unique ID:  101252125     Medline TA:  Clin Vaccine Immunol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  387-92     Citation Subset:  IM    
Affiliation:
Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, 1619 Campus Delivery, Fort Collins, CO 80523, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cats
Cells, Cultured
Clinical Laboratory Techniques / methods*
Immunoassay / methods
Interferon-gamma / analysis*,  secretion
Interleukin-10 / analysis*,  secretion
Interleukin-12 / analysis*,  secretion
Leukocytes, Mononuclear / immunology
Microspheres*
Sensitivity and Specificity
Grant Support
ID/Acronym/Agency:
5R01HL092791/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
130068-27-8/Interleukin-10; 187348-17-0/Interleukin-12; 82115-62-6/Interferon-gamma
Comments/Corrections

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