ABSTRACT
The development and evaluation of a Boc-AL(Boc)Q(Trt)-AMC fluorophore to detect 3C Protease, produced by Foot and Mouth Disease Virus (FMDV) is reported, with a view to a potential use as a rapid screen for FMDV infected livestock The peptide-linked conjugate fluorophore is evaluated in vitro for sensitivity, specificity, stability and rapidity and shows statistically significant increases in fluorescence when exposed to physiologically relevant concentrations of 3C Protease and selectivity when compared with other common proteases likely to be located, typically in the absence of FMDV. The stability of deprotected Boc-AL(Boc)Q(Trt)-AMC is reported as a limitation of this probe.
Subject(s)
Cysteine Endopeptidases/chemistry , Cysteine Endopeptidases/metabolism , Enzyme Assays/methods , Fluorescent Dyes/chemistry , Fluorescent Dyes/chemical synthesis , Foot-and-Mouth Disease Virus/enzymology , Viral Proteins/chemistry , Viral Proteins/metabolism , 3C Viral Proteases , Cell Line , Chemistry Techniques, SyntheticABSTRACT
This article reports the development of a novel fluorometric indicator which shows a rapid response when exposed to coagulase positive Staphylococcus aureus (SA) bacteria (including methicillin sensitive Staphylococcus aureus (MSSA) and methicillin resistant Staphylococcus aureus (MRSA) bacteria). The test is robust and will detect a wide variety of SA strains and there is no significant fluorescence response observed for other species of bacteria commonly found in clinical specimens, including other Staphylococcus bacteria. This research forms the basis of a prototype SA testing kit for the rapid detection of SA within hospital and healthcare environments as an economical prescreen or alternative to the current PCR based testing methodology. Rapid identification of SA carriers will allow hospital infection control teams to be pre-emptive and could significantly reduce the incidence of hospital acquired infections involving this organism.
