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

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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.

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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.

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Authors and Affiliations

  1. Department of Chemical Science and Technologies, Tor Vergata University, Via della Ricerca Scientifica, 00133, Rome, Italy

    Gianluca Adornetto, Laura Fabiani, Giulia Volpe, Alessia De Stefano & Danila Moscone

  2. Department of Pediatrics, La Sapienza University, Piazzale Aldo Moro 5, 00185, Rome, Italy

    Raffaella Nenna, Federica Lucantoni & Margherita Bonamico

  3. Radim SpA, Pomezia, Via del Mare, 00040, Pomezia, RM, Italy

    Sonia Martini

  4. Department of Internal Medicine and Medical Specialities, La Sapienza University, Via del Policlinico, 00161, Rome, Italy

    Claudio Tiberti

Authors
  1. Gianluca Adornetto
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  2. Laura Fabiani
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  3. Giulia Volpe
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  4. Alessia De Stefano
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  5. Sonia Martini
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  6. Raffaella Nenna
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  7. Federica Lucantoni
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  8. Margherita Bonamico
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  9. Claudio Tiberti
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  10. Danila Moscone
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Corresponding author

Correspondence to Danila Moscone.

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Adornetto, G., Fabiani, L., Volpe, G. et al. An electrochemical immunoassay for the screening of celiac disease in saliva samples. Anal Bioanal Chem 407, 7189–7196 (2015). https://doi.org/10.1007/s00216-015-8884-y

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  • DOI: https://doi.org/10.1007/s00216-015-8884-y

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