Multiplexed immunoassay for detection of rheumatoid factors by FIDISTM technology.
Our objective was to (i) compare FIDIS Rheuma, a new multiplexed immunoassay designed for simultaneous detection of IgM class rheumatoid factors (RF) directed against Fc determinants of IgG from humans and animals, with agglutination and ELISA (conventional methods) and (ii) evaluate the clinical sensitivity and specificity of biological markers for rheumatoid arthritis (RA). To do this, FIDIS technology was employed using the Luminex system. It consists of distinct color-coded microsphere sets, a flow cytometer, and digital signal processing hardware and software. Agglutination and ELISA tests were performed with commercial kits. The study included 134 samples from RA patients and 105 from healthy blood donors. For human specificity, we compared FIDIS with latex agglutination and ELISA. Relative sensitivities were 98.9% and 88.5% and specificities were 90.2% and 94.6%, respectively. For animal specificity, we compared FIDIS with Waaler-Rose and ELISA. The results were 84.9% and 71.9% for the sensitivities and 97.5% and 98.4% for the specificities, respectively. Detection of IgG anti-CCP by ELISA and IgG antikeratin by immunofluorescence was also determined in order to compare their clinical sensitivity and specificity with IgM-RF, according to the method used. The results were: IgG anti-CCP 72.3%, 97.2%; IgG antikeratin 36.6%, 100%; latex agglutination 66.4%, 97.2%; Waaler-Rose 55.9%, 96.3%; FIDIS human 73.9%, 92.1%; FIDIS animal 49.2%, 97.2%; ELISA human 93.2%, 95.5%; and ELISA animal 74.6%, 91.3%. The results showed the efficiency of FIDIS with analytical performance equivalent to the conventional methods, but having the advantage of giving quantitative results (IU/mL).
Abreu
I
I
Faculty of Medical Sciences, Department of Immunology, New University of Lisbon, Lisbon, Portugal. iabreu.imuno@fcm.unl.pt
Laroche
P
P
Bastos
A
A
Issert
V
V
Cruz
M
M
Nero
P
P
Fonseca
J E
JE
Branco
J
J
Machado Caetano
J A
JA
D000372:Agglutination Tests; D000818:Animals; D000918:Antibody Specificity; D001172:Arthritis, Rheumatoid / Q000175:diagnosis / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D016022:Case-Control Studies; D002956:Citrulline / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005069:Evaluation Studies as Topic; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007633:Keratins / Q000276:immunology; D007841:Latex Fixation Tests; D010456:Peptides, Cyclic / Q000276:immunology; D011817:Rabbits; D011933:Reagent Kits, Diagnostic; D012217:Rheumatoid Factor / Q000032:analysis*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D015415:Biomarkers; D000939:Epitopes; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D010456:Peptides, Cyclic; D011933:Reagent Kits, Diagnostic; D002956:Citrulline; D007633:Keratins; D012217:Rheumatoid Factor