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Analytical and Bioanalytical Chemistry

, Volume 403, Issue 4, pp 1191–1194 | Cite as

An ELIME assay for the rapid diagnosis of coeliac disease

  • Gianluca Adornetto
  • Giulia Volpe
  • Alessia De Stefano
  • Sonia Martini
  • Giuseppina Gallucci
  • Angelo Manzoni
  • Sergio Bernardini
  • Marco Mascini
  • Danila Moscone
Technical Note

Abstract

Coeliac disease (CD) is a gluten-induced autoimmune enteropathy found in genetically susceptible subjects. Because of the high number of undetected cases, rapid and cheaper screening methods are needed. Currently, the CD diagnosis involves the detection of anti-transglutaminase IgA antibodies (anti-tTG IgA) in blood serum through the use of ELISA systems with confirmation by histology of the intestinal mucosa. A new, rapid magneto-electrochemical immunosensor for CD diagnosis has been developed and applied to serum sample analysis. The system uses magnetic beads coated with tTG antigen to detect anti-tTG antibodies in positive serum samples and an alkaline phosphatase-conjugated anti-human IgA as label. An electrochemical readout, using magnetized screen-printed electrodes coupled with a portable instrument, is made after the addition of α-naphtyl phosphate, which is enzymatically converted into the electrochemically active α-naphthol product. The work involved the following considerations: (1) optimization of analytical parameters; (2) recovery evaluation, adding known concentrations of anti-tTG IgA to “blank” sera; (3) analysis of 107 blood serum samples; (4) calculation of the ROC curve, resulting in a cut-off of 1.0 U/ml, 100% of clinical sensitivity and 98.36% of clinical specificity; evaluation of the agreement between electrochemical and ELISA kit values (r 2 of 0.943). The system developed could be an useful tool for a correct and rapid CD diagnosis. This method is simple, cheap, rapid, and suitable for screening analyses performed outside of the classical diagnostic laboratory.

Keywords

Coeliac disease Transglutaminase Magnetic beads Electrochemical sensor 

Notes

Acknowledgments

The project, titled “Biosensori per la determinazione rapida (POCT) della celiachia e delle allergie.” has been funded by the Regione Lazio, in the Bando FILAS: Progetti di Ricerca Industriale e/o Sviluppo Sperimentale.

Supplementary material

216_2011_5702_MOESM1_ESM.pdf (849 kb)
ESM 1 (PDF 848 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Gianluca Adornetto
    • 1
  • Giulia Volpe
    • 1
  • Alessia De Stefano
    • 1
  • Sonia Martini
    • 2
  • Giuseppina Gallucci
    • 2
  • Angelo Manzoni
    • 3
  • Sergio Bernardini
    • 4
  • Marco Mascini
    • 5
  • Danila Moscone
    • 1
  1. 1.Dipartimento di Scienze e Tecnologie ChimicheUniversità di Roma Tor VergataRomeItaly
  2. 2.Radim SpAPomeziaItaly
  3. 3.SEAC-RADIM Group—Research and Development DepartmentCalenzanoItaly
  4. 4.Dipartimento di Medicina InternaUniversità di Roma Tor VergataRomeItaly
  5. 5.Dipartimento di ChimicaSesto FiorentinoItaly

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