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Updated: May 31, 2026
Guided Protocol for Fecal Microbial Characterization by 16S rRNA-Amplicon Sequencing
Published on: March 19, 2018
Correlation of rRNA gene amplicon pyrosequencing and bacterial culture for microbial compositional analysis of faecal
Ó O'Sullivan1, M Coakley, B Lakshminarayanan
1Teagasc Food Research Centre, Moorepark, Fermoy, Co. Cork, Ireland.
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.
