Rapid identification of pathogens responsible for necrotizing fasciitis on an integrated microfluidic system

Abstract

Necrotic fasciitis (NF) is a particularly aggressive and serious infection of the fascia that can penetrate into the musculature and internal organs, resulting in death if not treated promptly. In this work, an integrated microfluidic system composed of micropumps, microvalves, and micromixers was used to automate the detection of pathogens associated with NF. The entire molecular diagnostic process, including bacteria isolation, lysis, nucleic acid amplification and optical detection steps, was enacted on this developed system. Mannose binding lectin coated magnetic beads were first used as probes to isolate all bacteria in a sample. In this work, polymerase chain reaction assays featuring primers specific to genes from each of four NF-causing bacteria (, , and methicillin-sensitive and resistant) were used to rapidly and exclusively verify the presence of the respective bacterial strains, and the limits of detection were experimentally found to be 11, 1960, 14, and 11 400 colony forming units/reaction, respectively; all values reflect improvement over ones reported in literature. This integrated microfluidic chip may then be valuable in expediting diagnosis and optimizing treatment options for those with NF; such diagnostic improvements could ideally diminish the need for amputation and even reduce the morality rate associated with this life-threatening illness.

Early diagnosis of necrotizing fasciitis: Imaging techniques and their combined application

“…Despite these diagnostic scoring systems, clinical experience has shown that the diagnosis of NF relying solely on the above serological indicators is still insufficient and does not focus on the pathogenetic characterization of the trauma; therefore, some researchers have developed an integrated microfluidic system consisting of micropumps, microvalves, and micromixers, 48 which allows for the rapid identification of four pathogens (Vibrio vulnificus, Aeromonas hydrophila, and methicillin‐sensitive and resistant Staphylococcus aureus) associated with NF (within 90 min). Then by combining other methods for early diagnosis of NF, the accuracy of early diagnosis of NF was improved.…”