Abstract by Robert Hanson
Chemistry and Biochemistry
Combining Sequence-Specific DNA Hybridization with Single-Molecule Detection to improve Treatment of Sepsis
The treatment of acute sepsis induced by bacterial infection is often further complicated by the presence of drug resistance genes. Most cases require decisions on therapy to be made within two hours of a patient's admission, long before current diagnostic procedures can provide definitive information on the bacteria's drug resistance. This gap has led to survival rates as low as 50% in the case of some carbapenem resistant Enterobacteriaceae. We are working to develop diagnostics that can provide information on the drug resistance profile of bacteria related to sepsis within a 1 hour. We have created a microfluidic system capable of capturing specific drug resistance genes from fragmented plasmids isolated from whole blood. By hybridization to complementary strands attached to solid surfaces1, we have concentrated target gene sequences from 100 µL of a 500 fM samples. The DNA was then labeled with hybridization probes and detected on chip. We have developed methods for removal of the captured and labeled target DNA for analysis in a single-molecule detection system. These results are a key step toward creating an integrated system for diagnosing drug resistance.