Development of targeted genotyping methods for improved drug resistance surveillance [mPLEXABR]

Last updated on 3-12-2018 by Aurélie Felice
June 1, 2014
Project with no end date

In short

Countless genetic mechanisms can be responsible for the generation of bacteria resistant to antibiotics. For a detailed surveillance of drug resistance, it is paramount to identity the gene(s) causing the resistance. Some of these genes can be located on mobile elements, which can jump between bacteria, and thus rapidly spread drug resistance.

In this project, we aim to develop so-called ‘multiplex’ screens to identify the exact mechanism behind an observed resistance. This targeted approach is applicable to bacteria from either animal, food and human sources, allowing to map the circulation of antibiotics resistance genes countrywide and sector wide.

Project summary

The emergence of antibiotic resistant (ABR) bacteria is a serious public health concern. Such bacteria emerged following the intensive use of antimicrobials in humans and food-producing animals. Travel and intensive animal trade between EU member states promote the spread.

Public health laboratories perform surveillance of the precise types of circulating drug resistance genes. Their presence and spread is followed by systematically characterising the resistance profile of commensal and pathogenic bacteria isolated in different contexts.

Future surveillance programs would greatly profit from more detailed genetic insights into antibiotic resistance.

Current genetic characterisation is PCR-based and can only type one gene at the time. We can unravel the entire genome sequence of the pathogen, but this is still very costly. Therefore, there is a clear need for a cost-effective assay with broad scope, easy-to-use yet highly sensitive system which can compete with whole-genome sequencing technology as long as the latter remains unaffordable for routine use.

We are developing and validating low-cost (<5€/sample) genotyping assays for:

  1. β-lactamases
  2. fluoroquinolone resistance mechanisms in Salmonella and Shigella
  3. plasmid typing.

The presented method is based on a modular Multiplex Oligonucleotide Ligation-PCR procedure (MOL-PCR) and uses commercially available MagPlexTM-TAG microspheres beads for detection.

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