Abstract
Clostridium difficile is the primary cause of antibiotic associated diarrhea in humans and is a significant cause of morbidity and mortality. Thus the rapid and accurate identification of this pathogen in clinical samples, such as feces, is a key step in reducing the devastating impact of this disease. The bacterium produces two toxins, A and B, which are thought to be responsible for the majority of the pathology associated with the disease, although the relative contribution of each is currently a subject of debate. For this reason we have developed a rapid detection assay based on microwave-accelerated metal-enhanced fluorescence which is capable of detecting the presence of 10 bacteria in unprocessed human feces within 40 seconds. These promising results suggest that this prototype biosensor has the potential to be developed into a rapid, point of care, real time diagnostic assay for C. difficile.
Highlights
Clostridium difficile is a spore forming, toxin-producing bacterium which is currently the principal cause of healthcare associated diarrhea in the western world
The probes did not bind to bacterial species unrelated to C. difficile, further indicating that the probes were highly specific to toxins A and B of C. difficile
Species of the Clostridium genus, including species closely related to C. difficile did not show any probe hybridization which further validates probe specificity
Summary
Clostridium difficile is a spore forming, toxin-producing bacterium which is currently the principal cause of healthcare associated diarrhea in the western world. The bacterium currently presents a considerable challenge to healthcare professionals and has stimulated researchers to develop improved diagnostics and medical countermeasures. In healthy individuals the spore form of the bacterium is carried in the gut with no ill effects. Subsequent disruption of the gut flora, usually by broad spectrum antibiotics, enables the bacterium to proliferate and release two toxins, a 308 kDa enterotoxin (toxin A) and a 270 kDa cytotoxin (toxin B) which are responsible for symptoms ranging from mild diarrhea to fatal colitis [3]. The relative contribution of each toxin to pathogenicity is currently a subject of debate and the ability to detect the presence of both would greatly enhance diagnostic capability [7,8]
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