Article Text

other Versions

PDF
Examining the potential use and long-term stability of guaiac faecal occult blood test cards for microbial DNA 16S rRNA sequencing
  1. Morag Taylor1,
  2. Henry M Wood1,
  3. Stephen P Halloran2,
  4. Philip Quirke1
  1. 1Section of Pathology and Tumor Biology, Leeds Institute of Cancer and Pathology, University of Leeds, Leeds, UK
  2. 2NHS Bowel Cancer Screening Programme Hub, University of Surrey, Guildford, UK
  1. Correspondence to Morag Taylor, Section of Pathology and Tumour Biology, Level 4, Wellcome Trust Brenner Building, St James's University Hospital, Beckett Street, Leeds LS9 7TF, UK; medmta{at}leeds.ac.uk

Abstract

Aims With a growing interest in the influence the gut microbiome has on the development of colorectal cancer (CRC), we investigated the feasibility and stability of isolating and typing microbial DNA from guaiac faecal occult blood test (gFOBt) cards. This has the future potential to screen the microbial populations present in confirmed colorectal neoplasia cases with aims to predict the presence and development of CRC.

Methods Fresh stool samples from three healthy volunteers were applied to gFOBt cards. DNA was extracted from both the cards and fresh stool samples. A series of additional cards were prepared from one volunteer, and extracted at time points between 2 weeks and 3 years. The V4 region of the 16S rRNA gene was amplified and sequenced on an Illumina MiSeq at 2×250 bp read lengths. Data were analysed using QIIME software.

Results Samples were grouped both by volunteer and by type (fresh or gFOBt), and compared a variety of ways: visual inspection of taxa, α and β diversity, intraclass correlation. In all comparisons, samples grouped by volunteer, and not by sample type. The different time points showed no appreciable differences with increased storage time.

Conclusions This study has demonstrated that there is good concordance between microbial DNA isolated from fresh stool sample, and from the matched gFOBt card. Samples stored for up to 3 years showed no detrimental effect on measureable microbial DNA. This study has important future implications for investigating microbial influence on CRC development and other pathologies.

  • DNA
  • diagnostic screening
  • COLORECTAL CANCER
  • MICROBIOLOGY

Statistics from Altmetric.com

Footnotes

  • MT and HMW are joint first authors and have contributed equally.

  • Handling editor Runjan Chetty

  • Contributors MT: study concept and design, technical work and drafting of the manuscript. HMW: acquisition of data, data analysis and interpretation, drafting of the manuscript and obtaining funding. SPH: supply of faecal occult blood test cards, drafting manuscript and advice. PQ: Study concept and design, drafting of the manuscript and obtained funding.

  • Funding Laboratory consumables supported by a Cancer Research UK development fund. MT, HMW and PQ supported by Yorkshire Cancer Research. PQ is an NIHR Senior Investigator.

  • Competing interests None.

  • Provenance and peer review Not commissioned; internally peer reviewed.

  • Data sharing statement Raw sequence data are deposited in the European Nucleotide Archive, http://www.ebi.ac.uk/ena/data/view/PRJEB14174.

Request permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.