Brief description of project Goals: A miniature system enabling rapid, multiplexed, and portable point-of-care identification of antibiotic-resistant bacteria in under 15 minutes will be developed.
The rapid growth of antibiotic resistance is recognized as one of the most serious threats to public health in the U.S. and worldwide, with increasing numbers of drug-resistant bacteria leading to poor clinical outcomes and higher death rates, while consuming greater health-care resources. The ability to rapidly identify multiple bacteria and evaluate their antibiotic susceptibility at or near the point of care would greatly enhance the ability of clinicians to optimize treatment through the prescription of appropriate drugs based on the specific infections, or through the isolation of high risk patients to prevent further transmission. In this project we will develop a new technology using disposable lab-on-a-chip devices capable of segregating a patient sample into over 1000 individual reaction chambers where different microbial gene fragments associated with antibiotic resistance may be rapidly analyzed, allowing highly effective screening of multiple potential targets in under 15 min. The resulting technology is envisioned as a simple and inexpensive diagnostic platform for use at the point of care. The underlying lab-on-a-chip technologies developed in this work will also serve to advance other diagnostic platforms requiring simple operation and minimal equipment for use in clinical settings lacking central laboratory resources.
Antibiotic resistance represents a major and growing threat to public health, with drug-resistant pathogens significantly increasing rates of morbidity and mortality for infected patients. A major challenge associated with the increase in antimicrobial drug resistance is the lack of rapid assays for identifying causative pathogens and their drug resistance profiles during the earliest stages of treatment. In this project we will develop a low-cost and disposable thermoplastic microfluidic platform employing a novel trap array technology expected to open the door to routine clinical application of polymerase chain reaction (PCR) for antibiotic resistance screening at the point of care. The trap array platform will support over 1000 simultaneous PCR reactions using primers for multiple antibiotic-resistance gene targets, without the need for external pumping, valving, substrate preparation, or reagent introduction. The resulting platform will specifically be applied to the development of a rapid multiplexed assay for multi-drug resistant carbapenemase-producing bacteria, which currently presents a significant challenge across multiple clinical environments. To achieve these goals, the effort will result in the development of new technologies enabling (1) single-step discretization of a continuous sample volume into an array of individual sample traps to enable highly parallel analysis, (2) integration of all required PCR reagents, including defined primer sets for multiplexed analysis, directly into the array of sample traps during chip manufacture, and (3) rapid PCR and high resolution melt analysis (HRMA) of the resulting amplicons in a disposable thermoplastic chip requiring only a single manual pipetting step for operation, vastly simplifying assay implementation for use in near-patient settings without access to a central biochemical laboratory.
