Quantitative real-time PCR analysis of bacterial biomarkers enable fast and accurate monitoring in inflammatory bowel disease

PeerJ. 2022 Oct 18:10:e14217. doi: 10.7717/peerj.14217. eCollection 2022.

Abstract

Inflammatory bowel diseases (IBD) affect millions of people worldwide with increasing incidence. Ulcerative colitis (UC) and Crohn's disease (CD) are the two most common IBDs. There is no definite cure for IBD, and response to treatment greatly vary among patients. Therefore, there is urgent need for biomarkers to monitor therapy efficacy, and disease prognosis. We aimed to test whether qPCR analysis of common candidate bacteria identified from a patient's individual fecal microbiome can be used as a fast and reliable personalized microbial biomarker for efficient monitoring of disease course in IBD. Next generation sequencing (NGS) of 16S rRNA gene region identified species level microbiota profiles for a subset of UC, CD, and control samples. Common high abundance bacterial species observed in all three groups, and reported to be associated with IBD are chosen as candidate marker species. These species, and total bacteria amount are quantified in all samples with qPCR. Relative abundance of anti-inflammatory, beneficial Faecalibacterium prausnitzii, Akkermansia muciniphila, and Streptococcus thermophilus was significantly lower in IBD compared to control samples. Moreover, the relative abundance of the examined common species was correlated with the severity of IBD disease. The variance in qPCR data was much lower compared to NGS data, and showed much higher statistical power for clinical utility. The qPCR analysis of target common bacterial species can be a powerful, cost and time efficient approach for monitoring disease status and identify better personalized treatment options for IBD patients.

Keywords: Crohn’s disease; Inflammatory bowel disease; Molecular biomarker; Quantitative real-time PCR; Ulcerative colitis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / genetics
  • Biomarkers
  • Colitis, Ulcerative* / diagnosis
  • Crohn Disease* / diagnosis
  • Humans
  • Inflammatory Bowel Diseases* / diagnosis
  • RNA, Ribosomal, 16S / genetics
  • Real-Time Polymerase Chain Reaction

Substances

  • RNA, Ribosomal, 16S
  • Biomarkers

Grants and funding

This work was supported by the Turkish Society of Gastroenterology, and the IZTECH Scientific Research Projects Committee (BAP) (project number IYTE0209). There was no additional external funding received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.