Setup of Quantitative PCR for Oral <i>Neisseria</i> spp. Evaluation in Celiac Disease Diagnosis

• Main Authors: Maria Valeria Esposito, Carmela Nardelli, Ilaria Granata, Chiara Pagliuca, Valeria D’Argenio, Ilaria Russo, Mario Rosario Guarracino, Paola Salvatore, Giovanna Del Vecchio Blanco, Carolina Ciacci, Lucia Sacchetti
• Format: Article
• Language: English
• Published: MDPI AG 2019-12-01
• Series: Diagnostics
• Subjects: celiac disease; oral microbiome; <i>neisseria flavescens</i>; diagnostic marker; qpcr
• Online Access: https://www.mdpi.com/2075-4418/10/1/12

Coeliac disease (CD) is a multifactorial autoimmune disorder and gut dysbiosis contributes to its pathogenesis. We previously profiled by 16S rRNA sequencing duodenal and oropharyngeal microbiomes in active CD (a-CD), gluten-free diet (GFD) patients, and controls (CO) and found significantly higher levels of <i>Neisseria</i> spp., with pro-inflammatory activities, in a-CD patients than in the other two groups. In this study, we developed a fast and simple qPCR-based method to evaluate the abundance of the oral <i>Neisseria</i> spp. and the diagnostic performances of the test in CD diagnosis. The <i>Neisseria</i> spp. abundances detected by quantitative PCR (qPCR) were: CO = 0.14, GFD = 0.15, a-CD = 2.08, showing a similar trend to those previously measured by next generation sequencing (NGS). In particular, <i>Neisseria</i> spp. values obtained by both methods were significantly higher (<i>p</i> &lt; 0.001) in a-CD than in the other two groups GFD and CO&#8212;the latter almost overlapping. We calculated by ROC curve analysis the threshold of 1.12 ng/&#956;L of <i>Neisseria</i> spp. to discriminate between CO+GFD and a-CD patients with 100% and 96.7% of diagnostic sensitivity and specificity, respectively. In conclusion, our data, if confirmed in other cohorts, suggest the q-PCR evaluation of oral <i>Neisseria</i> spp. could be a fast and simple method to assess CD-associated dysbiosis for diagnostic purposes.