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


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.


Coeliac disease Transglutaminase Magnetic beads Electrochemical sensor 



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)


  1. 1.
    Ciclitira PJ, King AL, Fraser JS (2001) AGA technical review on celiac sprue. American Gastroenterological Association. Gastroenterology 120:636–651CrossRefGoogle Scholar
  2. 2.
    Maki M, Mustalahti K, Kokkonen J, Kulmala P, Haapalahti M, Karttunen T, Ilonen J, Laurila K, Dahlbom I, Hansson T, Hopfl P, Knip M (2003) Prevalence of celiac disease among children in Finland. N Engl J Med 348:2517–2524CrossRefGoogle Scholar
  3. 3.
    Sollid L, Thorsby E (1993) HLA subsceptibility genes in celiac disease: genetic mapping and role in pathogenesis. Gastroenterology 105:910–922Google Scholar
  4. 4.
    Catassi C, Fabiani E (1997) The spectrum of coelic disease in children. Bailliere Clin Gastroenterol 11:485–507CrossRefGoogle Scholar
  5. 5.
    Collin P, Reunala T, Pukkala E, Laippala P, Keyrilainen O, Pasternak A (1994) Celiac disease: associated disorders and survival. Gut 35:1215–1218CrossRefGoogle Scholar
  6. 6.
    Sigurgeirsson B, Agnarsson BA, Lindelof B (1994) Risk of lymphoma in patients with dermatitis herpetiformis. BMJ 308:13–15CrossRefGoogle Scholar
  7. 7.
    Lewis HM, Reunala TL, Garioch JJ, Leonard JN, Fry JS, Collin P, Evans D, Fry L (1996) Protective effect of gluten-free diet against development of lymphoma in dermatitis herpetiformis. Br J Dermatol 135:363–367CrossRefGoogle Scholar
  8. 8.
    Walker-Smith JA, Guandalini S, Schmitz J, Shmerling DH, Visakorpi JK (1990) Revised criteria for diagnosis of coeliac disease. Report of working group of European Society of Paediatric Gastroenterology and Nutrition. Arch Dis Child 65:909–911CrossRefGoogle Scholar
  9. 9.
    Dieterich W, Ehnis T, Bauer M, Donner P, Volta U, Riecken EO, Schuppan D (1997) Identification of tissue transglutaminase as the autoantigen of celiac disease. Nat Med 3:797–801CrossRefGoogle Scholar
  10. 10.
    Blackwell PJ, Hill PG, Holmes GKT (2002) Autoantibodies to human tissue transglutaminase: superior predictors of coeliac disease. Scand J Gastroenterol 37:1282–1285CrossRefGoogle Scholar
  11. 11.
    Volta U, Molinaro M, de Franceschi L, Fratangelo D, Bianchi FB (1995) IgA anti-endomysial antibodies on human umbilical cord tissue for celiac disease screening. Save both money and monkey. Dig Dis Sci 40:1902–1905CrossRefGoogle Scholar
  12. 12.
    Bonamico M, Tiberti C, Picarelli A, Mariani P, Rossi D, Cipolletta E, Greco M, Di Tola M, Sabbatella L, Carabba B, Magliocca FM, Strisciuglio P, Di Mario U (2001) Radioimmunoassay to detect antitranglutaminase autoantibodies is the most sensitive and specific screening method for celiac disease. AMJ Gastroenterol 96:1536–1540CrossRefGoogle Scholar
  13. 13.
    Catassi C, Ratsch IM, Fabiani E, Rossini M, Coppa GV, Giorgi P, Bordicchia F, Candela F (1994) Coeliac disease in the year 2000: exploring the iceberg. Lancet 343:200–203CrossRefGoogle Scholar
  14. 14.
    Gehring AG, Crawford CG, Mazenko RS, Van Houten NJ, Brewster JD (1996) Enzyme-linked immunomagnetic electrochemical detection of Salmonella Typhimurium. J Immunol Methods 195:15–25CrossRefGoogle Scholar
  15. 15.
    Henderson AR (1993) Assessing test accuracy and its clinical consequences: a primer for receiver operating characteristic curve analysis. Ann Clin Biochem 30:521–539Google Scholar
  16. 16.
    Swets JA (1998) Measuring the accuracy of diagnostic system. Science 240:1285–1293CrossRefGoogle Scholar

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

Personalised recommendations