An ELIME assay for the rapid diagnosis of coeliac disease |
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Authors: | Adornetto Gianluca Volpe Giulia De Stefano Alessia Martini Sonia Gallucci Giuseppina Manzoni Angelo Bernardini Sergio Mascini Marco Moscone Danila |
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Institution: | (1) Dipartimento di Scienze e Tecnologie Chimiche, Universit? di Roma Tor Vergata, Via della Ricerca Scientifica, 00133 Rome, Italy;(2) Radim SpA, Via del Mare 125, 00040 Pomezia, RM, Italy;(3) SEAC-RADIM Group—Research and Development Department, Via di Prato 74, 50041 Calenzano, Florence, Italy;(4) Dipartimento di Medicina Interna, Universit? di Roma Tor Vergata, 00133 Rome, Italy;(5) Dipartimento di Chimica, Via della Lastruccia 3, 50019 Sesto Fiorentino, Italy; |
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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. |
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