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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 23, pp 7189–7196 | Cite as

An electrochemical immunoassay for the screening of celiac disease in saliva samples

  • Gianluca Adornetto
  • Laura Fabiani
  • Giulia Volpe
  • Alessia De Stefano
  • Sonia Martini
  • Raffaella Nenna
  • Federica Lucantoni
  • Margherita Bonamico
  • Claudio Tiberti
  • Danila Moscone
Research Paper

Abstract

A highly sensitive electrochemical immunoassay for the initial diagnosis of celiac disease (CD) in saliva samples that overcomes the problems related to its high viscosity and to the low concentration of anti-transglutaminase antigen (tTG) IgA in this medium has been developed for the first time. The system uses magnetic beads (MBs) covered with tTG, which reacts with the anti-tTG IgA antibodies present in positive saliva samples. An anti-human IgA, conjugated with alkaline phosphate (AP) enzyme, was used as the label and a strip of eight magnetized screen-printed electrodes as the electrochemical transducer. In particular, two different immunoassay approaches were optimized and blindly compared to analyze a large number of saliva samples, whose anti-tTG IgA levels were independently determined by the radioimmunoassay (RIA) method. The obtained results, expressed as Ab index, were used to perform a diagnostic test evaluation through the construction of receiver operating characteristic (ROC) curves. The approach, involving a pre-incubation between the anti-human IgA-AP and saliva samples prior to the addition of MBs-tTG, showed a cutoff of 0.022 with 95 % clinical sensitivity and 96 % clinical specificity. The area under the ROC curve is equal to 1, a result that classifies our test as “perfect.” This study demonstrates that it is possible to perform the screening of CD with a rapid, simple, inexpensive, and sensitive method able to detect anti-tTG antibodies in saliva samples, which are easily obtained by non-invasive techniques. This aspect is of fundamental importance to screen a large number of subjects, especially in the pediatric age.

Graphical Abstract

ELIME assay Plus

Keywords

Magnetic beads Eight-SPE strip Celiac disease Transglutaminase Salivary anti-tTG antibodies 

Notes

Acknowledgments

The project entitled “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_2015_8884_MOESM1_ESM.pdf (14 kb)
ESM 1 (PDF 13 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Gianluca Adornetto
    • 1
  • Laura Fabiani
    • 1
  • Giulia Volpe
    • 1
  • Alessia De Stefano
    • 1
  • Sonia Martini
    • 3
  • Raffaella Nenna
    • 2
  • Federica Lucantoni
    • 2
  • Margherita Bonamico
    • 2
  • Claudio Tiberti
    • 4
  • Danila Moscone
    • 1
  1. 1.Department of Chemical Science and TechnologiesTor Vergata UniversityRomeItaly
  2. 2.Department of PediatricsLa Sapienza UniversityRomeItaly
  3. 3.Radim SpA, PomeziaPomeziaItaly
  4. 4.Department of Internal Medicine and Medical SpecialitiesLa Sapienza UniversityRomeItaly

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