Comparison of brush and biopsy sampling methods of the ileal pouch for assessment of mucosa-associated microbiota of human subjects

Susan M. Huse, Vincent B. Young, Hilary G. Morrison, Dionysios A. Antonopoulos, John Kwon, Sushila Dalal, Rose Arrieta, Nathaniel A. Hubert, Lici Shen, Joseph H. Vineis, Jason C. Koval, Mitchell L. Sogin, Eugene B. Chang, Laura E. Raffals

Research output: Contribution to journalArticle

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Abstract

Background: Mucosal biopsy is the most common sampling technique used to assess microbial communities associated with the intestinal mucosa. Biopsies disrupt the epithelium and can be associated with complications such as bleeding. Biopsies sample a limited area of the mucosa, which can lead to potential sampling bias. In contrast to the mucosal biopsy, the mucosal brush technique is less invasive and provides greater mucosal coverage, and if it can provide equivalent microbial community data, it would be preferable to mucosal biopsies.Results: We compared microbial samples collected from the intestinal mucosa using either a cytology brush or mucosal biopsy forceps. We collected paired samples from patients with ulcerative colitis (UC) who had previously undergone colectomy and ileal pouch anal anastomosis (IPAA), and profiled the microbial communities of the samples by sequencing V4-V6 or V4-V5 16S rRNA-encoding gene amplicons. Comparisons of 177 taxa in 16 brush-biopsy sample pairs had a mean R2 of 0.94. We found no taxa that varied significantly between the brush and biopsy samples after adjusting for multiple comparisons (false discovery rate ≤0.05). We also tested the reproducibility of DNA amplification and sequencing in 25 replicate pairs and found negligible variation (mean R2 = 0.99). A qPCR analysis of the two methods showed that the relative yields of bacterial DNA to human DNA were several-fold higher in the brush samples than in the biopsies.Conclusions: Mucosal brushing is preferred to mucosal biopsy for sampling the epithelial-associated microbiota. Although both techniques provide similar assessments of the microbial community composition, the brush sampling method has relatively more bacterial to host DNA, covers a larger surface area, and is less traumatic to the epithelium than the mucosal biopsy.

Original languageEnglish (US)
Article number5
JournalMicrobiome
Volume2
Issue number1
DOIs
StatePublished - Oct 8 2014

Fingerprint

Colonic Pouches
Microbiota
Mucous Membrane
Biopsy
Intestinal Mucosa
Epithelium
Bacterial DNA
Colectomy
Selection Bias
DNA
DNA Sequence Analysis
Ulcerative Colitis
rRNA Genes
Surgical Instruments
Cell Biology

Keywords

  • Cytology brush
  • Microbial sampling
  • Microbiome
  • Microbiome methods
  • Mucosal biopsy
  • Mucosal brushing
  • Ulcerative colitis

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Huse, S. M., Young, V. B., Morrison, H. G., Antonopoulos, D. A., Kwon, J., Dalal, S., ... Raffals, L. E. (2014). Comparison of brush and biopsy sampling methods of the ileal pouch for assessment of mucosa-associated microbiota of human subjects. Microbiome, 2(1), [5]. https://doi.org/10.1186/2049-2618-2-5

Comparison of brush and biopsy sampling methods of the ileal pouch for assessment of mucosa-associated microbiota of human subjects. / Huse, Susan M.; Young, Vincent B.; Morrison, Hilary G.; Antonopoulos, Dionysios A.; Kwon, John; Dalal, Sushila; Arrieta, Rose; Hubert, Nathaniel A.; Shen, Lici; Vineis, Joseph H.; Koval, Jason C.; Sogin, Mitchell L.; Chang, Eugene B.; Raffals, Laura E.

In: Microbiome, Vol. 2, No. 1, 5, 08.10.2014.

Research output: Contribution to journalArticle

Huse, SM, Young, VB, Morrison, HG, Antonopoulos, DA, Kwon, J, Dalal, S, Arrieta, R, Hubert, NA, Shen, L, Vineis, JH, Koval, JC, Sogin, ML, Chang, EB & Raffals, LE 2014, 'Comparison of brush and biopsy sampling methods of the ileal pouch for assessment of mucosa-associated microbiota of human subjects', Microbiome, vol. 2, no. 1, 5. https://doi.org/10.1186/2049-2618-2-5
Huse, Susan M. ; Young, Vincent B. ; Morrison, Hilary G. ; Antonopoulos, Dionysios A. ; Kwon, John ; Dalal, Sushila ; Arrieta, Rose ; Hubert, Nathaniel A. ; Shen, Lici ; Vineis, Joseph H. ; Koval, Jason C. ; Sogin, Mitchell L. ; Chang, Eugene B. ; Raffals, Laura E. / Comparison of brush and biopsy sampling methods of the ileal pouch for assessment of mucosa-associated microbiota of human subjects. In: Microbiome. 2014 ; Vol. 2, No. 1.
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abstract = "Background: Mucosal biopsy is the most common sampling technique used to assess microbial communities associated with the intestinal mucosa. Biopsies disrupt the epithelium and can be associated with complications such as bleeding. Biopsies sample a limited area of the mucosa, which can lead to potential sampling bias. In contrast to the mucosal biopsy, the mucosal brush technique is less invasive and provides greater mucosal coverage, and if it can provide equivalent microbial community data, it would be preferable to mucosal biopsies.Results: We compared microbial samples collected from the intestinal mucosa using either a cytology brush or mucosal biopsy forceps. We collected paired samples from patients with ulcerative colitis (UC) who had previously undergone colectomy and ileal pouch anal anastomosis (IPAA), and profiled the microbial communities of the samples by sequencing V4-V6 or V4-V5 16S rRNA-encoding gene amplicons. Comparisons of 177 taxa in 16 brush-biopsy sample pairs had a mean R2 of 0.94. We found no taxa that varied significantly between the brush and biopsy samples after adjusting for multiple comparisons (false discovery rate ≤0.05). We also tested the reproducibility of DNA amplification and sequencing in 25 replicate pairs and found negligible variation (mean R2 = 0.99). A qPCR analysis of the two methods showed that the relative yields of bacterial DNA to human DNA were several-fold higher in the brush samples than in the biopsies.Conclusions: Mucosal brushing is preferred to mucosal biopsy for sampling the epithelial-associated microbiota. Although both techniques provide similar assessments of the microbial community composition, the brush sampling method has relatively more bacterial to host DNA, covers a larger surface area, and is less traumatic to the epithelium than the mucosal biopsy.",
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AU - Morrison, Hilary G.

AU - Antonopoulos, Dionysios A.

AU - Kwon, John

AU - Dalal, Sushila

AU - Arrieta, Rose

AU - Hubert, Nathaniel A.

AU - Shen, Lici

AU - Vineis, Joseph H.

AU - Koval, Jason C.

AU - Sogin, Mitchell L.

AU - Chang, Eugene B.

AU - Raffals, Laura E.

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N2 - Background: Mucosal biopsy is the most common sampling technique used to assess microbial communities associated with the intestinal mucosa. Biopsies disrupt the epithelium and can be associated with complications such as bleeding. Biopsies sample a limited area of the mucosa, which can lead to potential sampling bias. In contrast to the mucosal biopsy, the mucosal brush technique is less invasive and provides greater mucosal coverage, and if it can provide equivalent microbial community data, it would be preferable to mucosal biopsies.Results: We compared microbial samples collected from the intestinal mucosa using either a cytology brush or mucosal biopsy forceps. We collected paired samples from patients with ulcerative colitis (UC) who had previously undergone colectomy and ileal pouch anal anastomosis (IPAA), and profiled the microbial communities of the samples by sequencing V4-V6 or V4-V5 16S rRNA-encoding gene amplicons. Comparisons of 177 taxa in 16 brush-biopsy sample pairs had a mean R2 of 0.94. We found no taxa that varied significantly between the brush and biopsy samples after adjusting for multiple comparisons (false discovery rate ≤0.05). We also tested the reproducibility of DNA amplification and sequencing in 25 replicate pairs and found negligible variation (mean R2 = 0.99). A qPCR analysis of the two methods showed that the relative yields of bacterial DNA to human DNA were several-fold higher in the brush samples than in the biopsies.Conclusions: Mucosal brushing is preferred to mucosal biopsy for sampling the epithelial-associated microbiota. Although both techniques provide similar assessments of the microbial community composition, the brush sampling method has relatively more bacterial to host DNA, covers a larger surface area, and is less traumatic to the epithelium than the mucosal biopsy.

AB - Background: Mucosal biopsy is the most common sampling technique used to assess microbial communities associated with the intestinal mucosa. Biopsies disrupt the epithelium and can be associated with complications such as bleeding. Biopsies sample a limited area of the mucosa, which can lead to potential sampling bias. In contrast to the mucosal biopsy, the mucosal brush technique is less invasive and provides greater mucosal coverage, and if it can provide equivalent microbial community data, it would be preferable to mucosal biopsies.Results: We compared microbial samples collected from the intestinal mucosa using either a cytology brush or mucosal biopsy forceps. We collected paired samples from patients with ulcerative colitis (UC) who had previously undergone colectomy and ileal pouch anal anastomosis (IPAA), and profiled the microbial communities of the samples by sequencing V4-V6 or V4-V5 16S rRNA-encoding gene amplicons. Comparisons of 177 taxa in 16 brush-biopsy sample pairs had a mean R2 of 0.94. We found no taxa that varied significantly between the brush and biopsy samples after adjusting for multiple comparisons (false discovery rate ≤0.05). We also tested the reproducibility of DNA amplification and sequencing in 25 replicate pairs and found negligible variation (mean R2 = 0.99). A qPCR analysis of the two methods showed that the relative yields of bacterial DNA to human DNA were several-fold higher in the brush samples than in the biopsies.Conclusions: Mucosal brushing is preferred to mucosal biopsy for sampling the epithelial-associated microbiota. Although both techniques provide similar assessments of the microbial community composition, the brush sampling method has relatively more bacterial to host DNA, covers a larger surface area, and is less traumatic to the epithelium than the mucosal biopsy.

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