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Development of a novel rapid non-invasive screening test for coeliac disease
  1. V Baldasa,
  2. A Tommasinia,
  3. C Trevisiola,
  4. I Bertia,
  5. A Fasanob,
  6. D Sblatteroa,
  7. A Bradburyc,
  8. R Marzarid,
  9. G Barillarie,
  10. A Venturaa,
  11. T Nota
  1. aDepartment of Pediatrics IRCCS “Burlo Garofolo”, Trieste, Italy, bDivision of Pediatric Gastroenterology and Nutrition, University of Maryland at Baltimore, USA, cInternational School for Advanced Studies, Trieste, Italy, dDepartment of Biology, Trieste, Italy, eBlood Transfusion Service, City Hospital of Udine, Italy
  1. T Not, Dipartimento di Pediatria, Istituto per l'Infanzia “Burlo Garofolo”, Via dell'Istria 65/1, 34100 Trieste, Italy.not{at}burlo.trieste.it

Abstract

BACKGROUND Coeliac disease is one of the commonest underdiagnosed diseases in general practice. The autoantigen recognised by the sera of patients with coeliac disease has recently been identified as tissue transglutaminase.

AIMS We evaluated a simple non-invasive immunological dot blot assay for coeliac disease, suitable for use by the general physician in the ambulatory setting. The sensitivity and specificity of this dot blot assay based on recognition of recombinant human transglutaminase were compared with those of antiendomysial antibodies and an enzyme linked immunosorbent assay.

METHODS Serum samples were analysed from 64 healthy controls, 58 first degree relatives of coeliacs, 74 diseased controls, and 70 biopsy confirmed untreated patients with coeliac disease. Dot blot assay and enzyme linked immunosorbent assay were performed using recombinant human transglutaminase as antigen.

RESULTS The dot blot assay, which can be performed in 20 minutes, was positive in all 70 untreated coeliacs (sensitivity 100%). Among the three control groups, there were three false positive tests by dot blot (specificity 98%), all belonging to the group of healthy subjects. The antiendomysial antibodies test missed five untreated coeliac patients (sensitivity 93%) and was negative in all three control groups (specificity 100%). The specificity of the immunosorbent assay was 99% for IgA and 98% for IgG, while sensitivity was 93% for IgA, 47% for IgG, and 100% for IgA and IgG combined.

CONCLUSIONS The dot blot assay is highly accurate in detecting untreated subjects with coeliac disease and can be performed in the general physician's medical office during the course of a routine examination. This innovative test is a practical, reliable alternative to both the immunofluorescent based antiendomysial test and immunosorbent assay for detection of transglutaminase antibodies for the diagnosis of coeliac disease.

  • coeliac disease
  • dot blot immunobinding assay
  • recombinant human tissue transglutaminase
  • Abbreviations used in this paper

    AEA
    antiendomysium antibodies
    CD
    coeliac disease
    ELISA
    enzyme linked immunosorbent assay
    h-tTG
    human tissue transglutaminase
    ELISA
    enzyme linked immunosorbent assay
    TBS
    Tris buffered saline
    TTBS
    Tween 20 Tris buffered saline
    PBS
    phosphate buffered saline
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  • Abbreviations used in this paper

    AEA
    antiendomysium antibodies
    CD
    coeliac disease
    ELISA
    enzyme linked immunosorbent assay
    h-tTG
    human tissue transglutaminase
    ELISA
    enzyme linked immunosorbent assay
    TBS
    Tris buffered saline
    TTBS
    Tween 20 Tris buffered saline
    PBS
    phosphate buffered saline
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