Introduction: Serum tTG-IgA is the first-line test for coeliac disease (CeD), but symptom-based, clinician-initiated testing detects only a minority of cases. Interest in population-based screening is therefore growing, creating a need for scalable, non-invasive tests. Saliva testing is attractive, although antibody detection has historically been limited by low analyte abundance. We evaluated whether an ultrasensitive antibody-dependent agglutination PCR assay could detect salivary tTG-IgA and identify active CeD in adults.
Methods: Unstimulated saliva and matched serum were collected from adults with biopsy-proven CeD, including newly diagnosed active CeD and treated CeD on a gluten-free diet, and from healthy controls. Salivary and serum tTG-IgA were measured using the ADAP assay, which detects antibody-mediated agglutination of tTG-DNA conjugates by PCR. Serum was also assessed using standard chemiluminescent immunoassay (CLIA; Inova QUANTA Flash). Diagnostic accuracy for active CeD was evaluated by ROC analysis. The impact of saliva collection method and 48-hour room-temperature storage, simulating postal transport, was assessed.
Results: The cohort included 43 adults (19 active CeD, 12 treated CeD, 12 healthy controls). All active CeD participants were serum tTG-IgA-positive by standard serology. Treated participants had followed a gluten-free diet for a mean of 7.6 years; 10/12 were tTG-IgA-negative. For distinguishing active CeD from treated CeD and controls, salivary ADAP showed 95% sensitivity, 100% specificity, and an AUROC of 0.98. Salivary ADAP correlated strongly with serum ADAP, r=0.72, p<0.0001, while serum CLIA and serum ADAP showed moderate correlation, r=0.46, p=0.0081. Passive drool collected via funnel produced a median 1.96-fold higher salivary tTG-IgA level than absorbent cotton swab-based Salivette collection. Salivary tTG-IgA remained stable after 48-hour room-temperature storage.
Conclusions: Salivary tTG-IgA was detectable in active CeD using an ultrasensitive ADAP assay and showed high diagnostic accuracy. Passive collection improved tTG-IgA yield, and room-temperature stability supports remote, at-home sample collection. Larger validation studies are underway.