scholarly journals Determination of Cytokine Responses Using a Multiplexed Fluorescent Microsphere Immunoassay

2002 ◽  
Vol 118 (3) ◽  
pp. 346-353 ◽  
Author(s):  
Thomas B. Martins ◽  
Brian M. Pasi ◽  
Jerry W. Pickering ◽  
Troy D. Jaskowski ◽  
Christine M. Litwin ◽  
...  
Keyword(s):  
Fluorescent Microsphere ◽  
Cytokine Responses ◽  
Multiplexed Fluorescent Microsphere Immunoassay ◽  
Microsphere Immunoassay

scholarly journals Evaluation of Multiplexed Fluorescent Microsphere Immunoassay for Detection of Autoantibodies to Nuclear Antigens

2004 ◽  
Vol 11 (6) ◽  
pp. 1054-1059 ◽  
Author(s):  
Thomas B. Martins ◽  
Rufus Burlingame ◽  
Carlos A. von Mühlen ◽  
Troy D. Jaskowski ◽  
Christine M. Litwin ◽  
...  
Keyword(s):  
Lupus Erythematosus ◽  
Vascular Diseases ◽  
Disease Diagnosis ◽  
Specific Method ◽  
Serum Samples ◽  
Collagen Vascular Diseases ◽  
Fluorescent Microsphere ◽  
Nuclear Antigens ◽  
Multiplexed Fluorescent Microsphere Immunoassay ◽  
Microsphere Immunoassay

ABSTRACT Antibodies to extractable nuclear antigens (ENA) are found in a variety of collagen vascular diseases. Determining the individual specificities of these antibodies is extremely useful in establishing the disease diagnosis and in some cases the prognosis. With a multiplexed fluorescent microsphere immunoassay, reactivity to five of the most diagnostically useful ENA was measured in 249 serum samples, including samples from 56 patients previously documented to have systemic lupus erythematosus (SLE). Results of the multiplexed assay were compared to results from established ENA enzyme-linked immunosorbent assays (ELISAs), and the agreement, sensitivity, and specificity, respectively, for the five ENA evaluated were as follows: SSA, 99.1, 100.0, and 98.8%; SSB, 98.6, 88.9, and 99.5%; Sm, 97.6, 95.8, and 97.9%; RNP, 97.2, 92.7, and 98.8%; Scl-70, 93.6, 50.0, and 99.0%. In the 56 confirmed SLE patients, the frequency of significant concentrations of autoantibodies with the multiplexed assay was 21.4% for SSA, 7.1% for SSB, 10.7% for Sm, 32.1% for RNP, and 0% for Scl-70. The new flow cytometric bead-based multiplexed assay showed excellent correlation with the well-established single-analyte ELISA methods for four of five the ENA markers investigated in this study. The most notable discrepancies between the two assays were for the Scl-70 antigen, which was most often resolved in favor of the multiplexed assay. Our studies show that the multiplexed microsphere-based immunoassay is a sensitive and specific method for the detection and semiquantitation of ENA antibodies in human sera.

scholarly journals A Multiplexed Fluorescent Microsphere Immunoassay for Antibodies to Pneumococcal Capsular Polysaccharides

2002 ◽  
Vol 117 (4) ◽  
pp. 589-596 ◽  
Author(s):  
Jerry W. Pickering ◽  
Thomas B. Martins ◽  
Ryan W. Greer ◽  
M. Carl Schroder ◽  
Mark E. Astill ◽  
...  
Keyword(s):  
Capsular Polysaccharides ◽  
Fluorescent Microsphere ◽  
Multiplexed Fluorescent Microsphere Immunoassay ◽  
Microsphere Immunoassay

scholarly journals Heterophile Antibody Interference in a Multiplexed Fluorescent Microsphere Immunoassay for Quantitation of Cytokines in Human Serum

2004 ◽  
Vol 11 (2) ◽  
pp. 325-329 ◽  
Author(s):  
Thomas B. Martins ◽  
Brian M. Pasi ◽  
Christine M. Litwin ◽  
Harry R. Hill
Keyword(s):  
Human Serum ◽  
Biological Fluids ◽  
Internal Controls ◽  
Fluorescent Microsphere ◽  
Simultaneous Quantification ◽  
And Control ◽  
Multiplexed Fluorescent Microsphere Immunoassay ◽  
Microsphere Immunoassay ◽  
Assay Interference

ABSTRACT While modern immunoassays provide sensitive and specific means for the quantitation of cytokines in biological fluids, heterophile antibodies are still a well-recognized cause of interference in the measurement of cytokines in these assays. We have developed a multiplexed fluorescent microsphere immunoassay for the simultaneous quantification of 10 cytokines in only 75 μl of serum. During the development of this multiplexed assay, the amount of assay interference due to heterophile antibodies was also determined, and methods for detecting heterophile interference and minimizing its effect were incorporated into the assay. Heterophile antibodies resulted in significantly elevated cytokine values compared to those of normal blood bank samples. These falsely elevated values, and thus the components of the assay the heterophile antibodies were binding to, were identified through the use of internal controls. This information was then used to design assay-specific blockers and absorbents that were shown to significantly reduce falsely elevated cytokine values while not affecting the standard and control values. The fluorescent multiplexed microsphere-based immunoassay can be used to quantitate multiple cytokines from a single sample and should be a useful tool in furthering our understanding of the role of cytokines in disease processes.

Development of a lateral flow fluorescent microsphere immunoassay for the determination of sulfamethazine in milk

2013 ◽  
Vol 405 (21) ◽  
pp. 6783-6789 ◽  
Author(s):  
Rui Chen ◽  
Heng Li ◽  
Han Zhang ◽  
Suxia Zhang ◽  
Weimin Shi ◽  
...  
Keyword(s):  
Lateral Flow ◽  
Fluorescent Microsphere ◽  
Microsphere Immunoassay

scholarly journals Development and validation of a multiplex fluorescent microsphere immunoassay assay for detection of porcine cytokines

MethodsX ◽  
2019 ◽  
Vol 6 ◽  
pp. 1218-1227
Author(s):  
S.A. Hall ◽  
S.H. Ison ◽  
C. Owles ◽  
J. Coe ◽  
D.A. Sandercock ◽  
...  
Keyword(s):  
Fluorescent Microsphere ◽  
Development And Validation ◽  
Microsphere Immunoassay

scholarly journals Rhinovirus species and tonsillar immune responses

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Emilia Mikola ◽  
Oscar Palomares ◽  
Riitta Turunen ◽  
Matti Waris ◽  
Lotta E. Ivaska ◽  
...  
Keyword(s):  
Immune Responses ◽  
Respiratory Symptoms ◽  
Clinical Characteristics ◽  
Respiratory Viruses ◽  
Positive Group ◽  
Cytokine Responses ◽  
Tonsil Tissue ◽  
Ifn Γ ◽  
Rhinovirus C

Abstract Background Rhinovirus A and C infections are important contributors to asthma induction and exacerbations. No data exist on the interaction of local immune responses in rhinovirus infection. Therefore, we aimed to determine the tonsillar immune responses according to rhinovirus A, B and C infections. Methods We collected tonsillar samples, nasopharyngeal aspirates and peripheral blood from 42 rhinovirus positive tonsillectomy patients. Fifteen respiratory viruses or their types were investigated from nasopharynx and tonsil tissue, and rhinovirus species were typed. The expression of 10 cytokines and 4 transcription factors (IFN-α, IFN-β, IFN-γ, IL-10, IL-13, IL-17, IL-28, IL-29, IL-37, TGF-β, FOXP3, GATA3, RORC2 and Tbet) were studied from tonsil tissue by quantitative PCR. A standard questionnaire of respiratory symptoms and health was filled by the patient or his/her guardian. The patients were divided into three groups by the determination of rhinovirus species. Results Overall, 16 patients had rhinovirus A, 12 rhinovirus B and 14 rhinovirus C infection. In rhinovirus B positive group there were significantly less men (P = 0.0072), less operated in spring (P = 0.0096) and more operated in fall (P = 0.030) than in rhinovirus A or C groups. Rhinovirus A positive patients had more respiratory symptoms (P = 0.0074) and particularly rhinitis (P = 0.036) on the operation day. There were no significant differences between the groups in virus codetection. In adjusted analysis, rhinovirus C infections were associated with increased IFN-α (P = 0.045) and decreased RORC2 expression (P = 0.025). Conclusions Rhinovirus species associated differently with clinical characteristics and tonsillar cytokine responses.

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