LFRET, a novel rapid assay for anti-tissue transglutaminase antibody detection

Autoři: Juuso Rusanen aff001;  Anne Toivonen aff002;  Jussi Hepojoki aff001;  Satu Hepojoki aff001;  Pekka Arikoski aff005;  Markku Heikkinen aff006;  Outi Vaarala aff007;  Jorma Ilonen aff008;  Klaus Hedman aff001
Působiště autorů: University of Helsinki, Medicum, Department of Virology, Helsinki, Finland aff001;  Laboratory Services (HUSLAB), Department of Virology and Immunology, University of Helsinki and Helsinki University Hospital, Helsinki, Finland aff002;  Department of Clinical Microbiology, Institute of Clinical Medicine, University of Eastern Finland, Kuopio, Finland aff003;  Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland aff004;  Department of Pediatrics, Kuopio University Hospital and University of Eastern Finland, Kuopio, Finland aff005;  Department of Gastroenterology, Kuopio University Hospital, Kuopio, Finland aff006;  Clinicum, University of Helsinki, Helsinki, Finland aff007;  Immunogenetics Laboratory, Institute of Biomedicine, University of Turku and Clinical Microbiology, Turku University Hospital, Turku, Finland aff008
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0225851


The diagnosis of celiac disease (CD) is currently based on serology and intestinal biopsy, with detection of anti-tissue transglutaminase (tTG) IgA antibodies recommended as the first-line test. Emphasizing the increasing importance of serological testing, new guidelines and evidence suggest basing the diagnosis solely on serology without confirmatory biopsy. Enzyme immunoassays (EIAs) are the established approach for anti-tTG antibody detection, with the existing point-of-care (POC) tests lacking sensitivity and/or specificity. Improved POC methods could help reduce the underdiagnosis and diagnostic delay of CD. We have previously developed rapid homogenous immunoassays based on time-resolved Förster resonance energy transfer (TR-FRET), and demonstrated their suitability in serodiagnostics with hanta- and Zika virus infections as models. In this study, we set out to establish a protein L -based TR-FRET assay (LFRET) for the detection of anti-tTG antibodies. We studied 74 patients with biopsy-confirmed CD and 70 healthy controls, with 1) the new tTG-LFRET assay, and for reference 2) a well-established EIA and 3) an existing commercial POC test. IgG depletion was employed to differentiate between anti-tTG IgA and IgG positivity. The sensitivity and specificity of the first-generation tTG-LFRET POC assay in detection of CD were 87.8% and 94.3%, respectively, in line with those of the reference POC test. The sensitivity and specificity of EIA were 95.9% and 91.9%, respectively. This study demonstrates the applicability of LFRET to serological diagnosis of autoimmune diseases in general and of CD in particular.

Klíčová slova:

Autoimmune diseases – Biopsy – Diagnostic medicine – Energy transfer – Enzyme-linked immunoassays – Serodiagnosis – Celiac disease – Enzyme immunoassay


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Článek vyšel v časopise


2019 Číslo 11