Magneto immunofluorescence assay for diagnosis of celiac disease

Silvina V. Kergaravat, Luis Beltramino, Nidia Garnero, Liliana Trotta, Marta Wagener, Silvia N. Fabiano, Maria Isabel Pividori, Silvia R. Hernandez

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

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 8.1U 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. © 2013 Elsevier B.V.
Original languageEnglish
Pages (from-to)89-96
JournalAnalytica Chimica Acta
Volume798
DOIs
Publication statusPublished - 10 Oct 2013

Keywords

  • Anti-transglutaminase antibodies
  • Celiac disease
  • ELISA
  • Fluorescence response
  • Magnetic beads
  • Receiver-operating characteristic

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