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Quantification of class 1 integron abundance in natural environments using real-time quantitative PCR
Authors:Hardwick Simon A  Stokes H W  Findlay Sophia  Taylor Mark  Gillings Michael R
Institution:Department of Biological Sciences, Genes to Geosciences Research Centre, Macquarie University, Sydney, Australia;;Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia;and;Department of Physical Geography, Macquarie University, Sydney, Australia
Abstract:Integrons are bacterial genetic elements capable of capturing and expressing potentially adaptive genetic material. Class 1 integrons constitute the most intensely studied group of these elements to date, mainly due to their well-established role in the acquisition and dissemination of antibiotic resistance genes in clinical environments. However, virtually nothing is known about the distribution or abundance of class 1 integrons outside of the clinical context. Here we develop a SYBR Green-based real-time quantitative PCR assay capable of quantifying the abundance of class 1 integrons in environmental samples. It was shown that the abundance of the intI1 gene in creek sediment correlates with ecological condition, implying that class 1 integrons provide selective advantages relevant to environmental pressures other than the use of antibiotics. By comparing the quantities of intI1 and 16S rRNA gene in each sample, it was demonstrated that approximately 2.7% of cells potentially harbour a class 1 integron. These findings suggest that class 1 integrons are widespread in natural environments removed from clinical settings and occur in a broader range of host organisms than had previously been assumed on the basis of culture-dependent estimates.
Keywords:class 1 integron              intI1            real-time qPCR  SYBR Green  culture independent
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