Recent progress in disease research and molecular diagnostics are leading us to the realization stage of highly precise and personalized treatment technology. However, complicated sample preparation process and low precision of quantitative (q)PCR in current molecular diagnostic technology are enormously delaying the development of treatment for organisms which are causes of healthcare-associated infections, such as Enterobacteriaceae, E. coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Here we develop an integrated drug-resistant Gram-negative bacteria (GNB) detection on chip that allows highly selective capture, rapid bacterial lysis, and precise gene expression measurements. We integrate microfluidic components for highly selective capture, rapid bacterial lysis, and precise digital PCR in order to achieve a single-chip molecular diagnostic technologies without any contaminations and loss of low number target pathogens in clinical sample. The device can both highly selective immuno-capture and effective in-situ photothermally-activated thermal lysis of GNBs with four different types of antibody immobilized on gold coated micropillar array. After the on-chip selective capture and in-situ thermal lysis, the lysates go through a high-reliability digital single DNA amplification in 1,200,000 reaction chambers of picoliter volume with density of 80,000 chambers/cm2, which can achieve reduced contamination via uniform partitioning of single DNA molecules and effective detection with high sensitivity and precision. Precise and rapid on-chip integrated molecular diagnostic technology will greatly help us develop highly personalized medicines for predictive medical treatments.
Project end date: 01/26/16