Abstract by David Harris
Chemistry and Biochemistry
Optimizing conditions for the capture, enrichment, labeling and detection of DNA from antibiotic resistant genes in sepsis
Antibiotic resistant infections are a growing health care concern, with new cases being reported annually. These infections can cause irreversible harm or death without timely diagnosis. However, the current diagnostic tests require cell culture which takes at least 24 hours, or PCR which requires extra reagents and careful sample preparation. Our system bypasses the problems with current methods by capturing and enriching DNA using magnetic microbeads in microfluidic devices. The magnetic microbeads are modified with ssDNA complementary to target DNA from antibiotic resistance genes. After hybridization and pull down, all other material is removed, and the captured DNA is labeled by hybridization and detected. Presently, our limit of detection is ~1 nM which is too high to be effective in a clinical setting. We are testing different methods to improve capture efficiency in microfluidics, particularly electromagnetic mixing of beads.