| Development and validation of a multiplex microsphere-based assay for detection of domestic cat (Felis catus) cytokines. | |
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MedLine Citation:
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PMID: 21209158 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Britta A Wood; Kevin P O'Halloran; Sue Vandewoude |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural Date: 2011-01-05 |
Journal Detail:
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Title: Clinical and vaccine immunology : CVI Volume: 18 ISSN: 1556-679X ISO Abbreviation: Clin. Vaccine Immunol. Publication Date: 2011 Mar |
Date Detail:
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Created Date: 2011-03-03 Completed Date: 2011-06-07 Revised Date: 2011-09-13 |
Medline Journal Info:
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Nlm Unique ID: 101252125 Medline TA: Clin Vaccine Immunol Country: United States |
Other Details:
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Languages: eng Pagination: 387-92 Citation Subset: IM |
Affiliation:
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Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, 1619 Campus Delivery, Fort Collins, CO 80523, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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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:
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5R01HL092791/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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130068-27-8/Interleukin-10; 187348-17-0/Interleukin-12; 82115-62-6/Interferon-gamma |
| Comments/Corrections | |
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