Magneto immunofluorescence assay for diagnosis of celiac disease.
A magneto immunofluorescence assay for the detection of anti-transglutaminase antibodies (ATG2) in celiac disease was developed. The ATG2 were recognized by transglutaminase enzyme immobilized on the magnetic beads and then the immunological reaction was revealed by antibodies labeled with peroxidase. The fluorescent response of the enzymatic reaction with o-phenylenediamine and H2O2 as substrates was correlated with anti-transglutaminase titer, showing EC50 and LOD values of 1:11,600 and 1:74,500 of antibody titers, respectively. A total number of 29 sera samples from clinically confirmed cases of celiac disease and 19 negative control samples were tested by the novel magneto immunofluorescence assay. The data were submitted to the receiver-operating characteristic plot (ROC) analysis which indicated that 8.1 U was the most effective cut-off value to discriminate correctly between celiac and non-celiac patients. The immunofluorescence assay exhibited a sensitivity of 96.6%, a specificity of 89.5% and an efficiency 93.8% compared with the commercial optical ELISA kit.
Laboratorio de Sensores y Biosensores, Cátedra de Química Analítica I, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, 3000 Santa Fe, Argentina.
Silvina V
SV
Kergaravat
Luis
L
Beltramino
Nidia
N
Garnero
Liliana
L
Trotta
Marta
M
Wagener
Silvia N
SN
Fabiano
Maria Isabel
MI
Pividori
Silvia R
SR
Hernandez
D000906:Antibodies / Q000097:blood* / Q000276:immunology; D002446:Celiac Disease / Q000097:blood / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000276:immunology; D019084:Fluorescent Antibody Technique, Indirect; D019204:GTP-Binding Proteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D008280:Magnetics; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D012372:ROC Curve; D011503:Transglutaminases / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D004800:Enzymes, Immobilized; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins
Anti-transglutaminase antibodies; Celiac disease; ELISA; Fluorescence response; Magnetic beads; Receiver-operating characteristic