A degenerate PCR-based strategy as a means of identifying homologues of aminoglycoside and β-lactam resistance genes in the gut microbiota.
BACKGROUND
The potential for the human gut microbiota to serve as a reservoir for antibiotic resistance genes has been the subject of recent discussion. However, this has yet to be investigated using a rapid PCR-based approach. In light of this, here we aim to determine if degenerate PCR primers can detect aminoglycoside and β-lactam resistance genes in the gut microbiota of healthy adults, without the need for an initial culture-based screen for resistant isolates. In doing so, we would determine if the gut microbiota of healthy adults, lacking recent antibiotic exposure, is a reservoir for resistance genes.


RESULTS
The strategy employed resulted in the identification of numerous aminoglycoside (acetylation, adenylation and phosphorylation) and β-lactam (including blaOXA, blaTEM, blaSHV and blaCTX-M) resistance gene homologues. On the basis of homology, it would appear that these genes originated from different bacterial taxa, with members of the Enterobacteriaceae being a particularly rich source. The results demonstrate that, even in the absence of recent antibiotic exposure, the human gut microbiota is a considerable reservoir for antibiotic resistance genes.


CONCLUSIONS
This study has demonstrated that the gut can be a significant source of aminoglycoside and β-lactam resistance genes, even in the absence of recent antibiotic exposure. The results also demonstrate that PCR-based approaches can be successfully applied to detect antibiotic resistance genes in the human gut microbiota, without the need to isolate resistant strains. This approach could also be used to rapidly screen other complex environments for target genes.
Fiona
F
Fouhy
R Paul
RP
Ross
Gerald F
GF
Fitzgerald
Catherine
C
Stanton
Teagasc Food Research Centre, Moorepark, Fermoy, Cork, Ireland. paul.cotter@teagasc.ie.
Paul D
PD
Cotter
D000328:Adult; D000617:Aminoglycosides / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial; D004755:Enterobacteriaceae / Q000187:drug effects / Q000235:genetics*; D041981:Gastrointestinal Tract / Q000382:microbiology*; D005798:Genes, Bacterial; D064368:Healthy Volunteers; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D047090:beta-Lactams / Q000494:pharmacology*
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D047090:beta-Lactams