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Replicon sequence typing of IncF plasmids carrying virulence and resistance determinants

Laura Villa1, Aurora García-Fernández, Daniela Fortini

  • 1Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy.

The Journal of Antimicrobial Chemotherapy
|October 12, 2010
PubMed
Abstract

Insights

A new rapid sequence-based typing method effectively categorizes IncF plasmids, which spread antimicrobial resistance and virulence genes. This approach aids in understanding the distribution and evolution of these important clinical plasmids.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • IncF plasmids are prevalent in clinical enterobacteria, facilitating the spread of antimicrobial resistance and virulence genes.
  • Their heterogeneity and multiple replicons pose challenges for traditional typing methods like restriction fragment length polymorphism analysis.
  • Accurate characterization of IncF plasmids is crucial for tracking the dissemination of resistance and virulence factors.

Purpose of the Study:

  • To develop a rapid, sequence-based typing scheme for categorizing IncF plasmids into homogeneous groups.
  • To improve the discrimination and analysis of IncF plasmid variants, particularly those carrying significant resistance and virulence genes.
  • To enhance epidemiological surveillance and evolutionary studies of IncF plasmids.

Main Methods:

  • Compared available IncF replicon sequences to identify discriminating characteristics.
  • Developed a typing method based on PCR amplification and sequence typing of IncF replicons (Replicon Sequence Typing - RST).
  • Tested the RST system on a collection of IncF plasmids from diverse sources and geographical origins.

Main Results:

  • A novel Replicon Sequence Typing (RST) scheme was successfully devised to differentiate IncF plasmid variants.
  • The RST system effectively discriminated between IncF plasmids carrying the same resistance gene (e.g., bla(CTX-M-15)) and related virulence plasmids (e.g., IncFIme).
  • The PCR-based replicon typing (PBRT) system was updated with new primers for identifying IncF plasmids from Salmonella, Klebsiella, and Yersinia species.

Conclusions:

  • The developed RST scheme enables the recognition and sub-categorization of IncF plasmids into homogeneous groups based on phylogenetic relatedness.
  • This method facilitates the analysis of IncF plasmid distribution in various environments.
  • The RST approach aids in uncovering the evolutionary origins and dissemination pathways of clinically relevant IncF plasmids.

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