Setup of Quantitative PCR for Oral <i>Neisseria</i> spp. Evaluation in Celiac Disease Diagnosis
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
Affiliations
Maria Valeria Esposito
Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, 80131 Naples, Italy
Carmela Nardelli
Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, 80131 Naples, Italy
Ilaria Granata
LabGTP (Laboratory of Genomics, Transcriptomics and Proteomics), Institute for High Performance Computing and Networking (ICAR), National Research Council (CNR), 80131 Naples, Italy
Chiara Pagliuca
Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, 80131 Naples, Italy
Valeria D’Argenio
Ceinge Biotecnologie Avanzate S. C. a R. L., 80131 Naples, Italy
Ilaria Russo
Department of Medicine and Surgery, University of Salerno, 84084 Salerno, Italy
Mario Rosario Guarracino
LabGTP (Laboratory of Genomics, Transcriptomics and Proteomics), Institute for High Performance Computing and Networking (ICAR), National Research Council (CNR), 80131 Naples, Italy
Paola Salvatore
Department of Molecular Medicine and Medical Biotechnologies, University of Naples Federico II, 80131 Naples, Italy
Giovanna Del Vecchio Blanco
Department of System Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Carolina Ciacci
Department of Medicine and Surgery, University of Salerno, 84084 Salerno, Italy
Lucia Sacchetti
Ceinge Biotecnologie Avanzate S. C. a R. L., 80131 Naples, Italy
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 Neisseria 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 Neisseria spp. and the diagnostic performances of the test in CD diagnosis. The Neisseria 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, Neisseria spp. values obtained by both methods were significantly higher (p < 0.001) in a-CD than in the other two groups GFD and CO—the latter almost overlapping. We calculated by ROC curve analysis the threshold of 1.12 ng/μL of Neisseria 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 Neisseria spp. could be a fast and simple method to assess CD-associated dysbiosis for diagnostic purposes.
