Characterization and Expression of Drug Resistance Genes in MDROs Originating from Combat Wound Infections
Technical Report,13 Sep 2012,12 Jun 2016
J Craig Venter Institute Rockville United States
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Twenty-five multi-drug resistant organisms MDROs of high military importance were sequenced using the Illumina HiSeq platform to identify drug resistance and virulence mechanism contributing to their success as pathogens in the wound environment. A novel bioinformatics pipeline was used to incorporate genomic sequencing data obtained from multiple sequencing platforms with optical restriction map data to assemble the genomes into sequence contigs. Bioinformatics processes for the automated identification and classification of antibiotic resistance mechanisms and IS elements were developed to enable mapping of these features onto the genomes of the 25 selected MDROs. Preliminary investigations of IS element content showed considerable diversity and abundance levels across a collection of Acinetobacter genomes. Three top priority MDROs and three commensal microbes were chosen and obtained from WRAIR MRSN for in vitro co-culturing in confrontation assays aimed at investigating alterations in gene expression, particularly antibiotic resistance genes and virulence factors. Mock confrontations using E. coli and S. aureus controls have been conducted for the development of a new solid-media based confluent lawn co-culturing assay, and an RNA extraction protocol that is biologically reproducible. Human cell cultures to be used in bacterial-human cell confrontations have been received and grown successfully in the lab.
- Genetic Engineering and Molecular Biology