Cargando…
Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis
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...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7168164/ https://www.ncbi.nlm.nih.gov/pubmed/31888008 http://dx.doi.org/10.3390/diagnostics10010012 |
_version_ | 1783523624416706560 |
---|---|
author | Esposito, Maria Valeria Nardelli, Carmela Granata, Ilaria Pagliuca, Chiara D’Argenio, Valeria Russo, Ilaria Guarracino, Mario Rosario Salvatore, Paola Del Vecchio Blanco, Giovanna Ciacci, Carolina Sacchetti, Lucia |
author_facet | Esposito, Maria Valeria Nardelli, Carmela Granata, Ilaria Pagliuca, Chiara D’Argenio, Valeria Russo, Ilaria Guarracino, Mario Rosario Salvatore, Paola Del Vecchio Blanco, Giovanna Ciacci, Carolina Sacchetti, Lucia |
author_sort | Esposito, Maria Valeria |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7168164 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-71681642020-04-21 Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis Esposito, Maria Valeria Nardelli, Carmela Granata, Ilaria Pagliuca, Chiara D’Argenio, Valeria Russo, Ilaria Guarracino, Mario Rosario Salvatore, Paola Del Vecchio Blanco, Giovanna Ciacci, Carolina Sacchetti, Lucia Diagnostics (Basel) Communication 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. MDPI 2019-12-26 /pmc/articles/PMC7168164/ /pubmed/31888008 http://dx.doi.org/10.3390/diagnostics10010012 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Communication Esposito, Maria Valeria Nardelli, Carmela Granata, Ilaria Pagliuca, Chiara D’Argenio, Valeria Russo, Ilaria Guarracino, Mario Rosario Salvatore, Paola Del Vecchio Blanco, Giovanna Ciacci, Carolina Sacchetti, Lucia Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis |
title | Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis |
title_full | Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis |
title_fullStr | Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis |
title_full_unstemmed | Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis |
title_short | Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis |
title_sort | setup of quantitative pcr for oral neisseria spp. evaluation in celiac disease diagnosis |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7168164/ https://www.ncbi.nlm.nih.gov/pubmed/31888008 http://dx.doi.org/10.3390/diagnostics10010012 |
work_keys_str_mv | AT espositomariavaleria setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT nardellicarmela setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT granatailaria setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT pagliucachiara setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT dargeniovaleria setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT russoilaria setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT guarracinomariorosario setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT salvatorepaola setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT delvecchioblancogiovanna setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT ciaccicarolina setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis AT sacchettilucia setupofquantitativepcrfororalneisseriasppevaluationinceliacdiseasediagnosis |