Correlation of rRNA gene amplicon pyrosequencing and bacterial culture for microbial compositional analysis of faecal samples from elderly Irish subjects

J Appl Microbiol. 2011 Aug;111(2):467-73. doi: 10.1111/j.1365-2672.2011.05067.x. Epub 2011 Jul 1.

Abstract

Aims: The aim of this investigation was to establish the degree of correlation between measurements from culture-dependent microbiological techniques and from next generation sequencing technologies.

Methods and results: Data generated by both techniques were collected from faecal samples from 185 elderly Irish people involved in the ongoing ELDERMET study (http://eldermet.ucc.ie). The results for three groups of intestinal bacteria were compared. Bifidobacterium sp., Lactobacillus sp. and Enterobacteriaceae were enumerated on selective media through culture-dependent techniques, whereas proportions of these bacteria were determined through sequencing technology against the background of other bacteria. The Spearman's rank correlation coefficient determined a good correlation between results from culture-dependent microbiology and culture-independent techniques for all three bacterial groups assessed (correlation coefficients for Bifidobacterium sp., Lactobacillus sp. and Enterobacteriaceae were 0·380, 0·366 and 0·437, respectively).

Conclusion: Correlation between the two methods implies that a single method is capable of profiling intestinal Bifidobacterium, Lactobacillus and Enterobacteriaceae populations. However, both methods have advantages that justify their use in tandem.

Significance and impact of the study: This is the first extensive study to compare bacterial counts from culture-dependent microbiological techniques and from next generation sequencing technologies.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Bifidobacterium / genetics
  • Bifidobacterium / isolation & purification*
  • Colony Count, Microbial
  • Computational Biology
  • Culture Media
  • DNA, Bacterial / genetics
  • Enterobacteriaceae / genetics
  • Enterobacteriaceae / isolation & purification*
  • Feces / microbiology*
  • Genes, rRNA
  • Humans
  • Intestines / microbiology*
  • Lactobacillus / genetics
  • Lactobacillus / isolation & purification*
  • Sequence Analysis, DNA

Substances

  • Culture Media
  • DNA, Bacterial