Key result
A fluorescent labeling strategy using Quantum dots bound to biotinylated capsids yielded a relative fluorescence of 528 ± 48.1 for bio-MNV-1 compared to 112 ± 8.6 for control MNV-1.
Why the study?
Studying interactions between enteric pathogens and their environment is challenging due to difficulties in tracking pathogens in their natural environments.
Absolute Event Rate: 528% vs 112%
A novel fluorescent labeling strategy using Quantum dots allows for in situ visualization of enteric viruses without affecting their infectivity.
Enables virus tracking in animal models; leaves open translation to human enteric virus research.
Studying the interactions between enteric pathogens and their environment is important to improving our understanding of their persistence and transmission. However, this remains challenging in large part because of difficulties associated with tracking pathogens in their natural environment(s). In this study, we report a fluorescent labeling strategy which was applied to murine norovirus (MNV-1), a human norovirus surrogate, and hepatitis A virus (HAV). Specifically, streptavidin-labeled Quantum dots (Q-Dots) were bound to biotinylated capsids of MNV-1 and HAV (bio-MNV-1 and bio-HAV); the process was confirmed by using a sandwich-type approach in which streptavidin-bound plates were reacted with biotinylated virus followed by a secondary binding to Q-Dots with an emission range of 635 to 675 nm (Q-Dots 655). The assay demonstrated a relative fluorescence of 528 +/- 48.1 and 112 +/- 8.6 for bio-MNV-1 and control MNV-1, respectively. The biotinylation process did not impact virus infectivity, nor did it interfere with the interactions between the virus and host cells or model produce items. Using fluorescent microscopy, it was possible to visualize both bio-HAV and bio-MNV-1 attached to the surfaces of permissive mammalian cells and green onion tissue. The method provides a powerful tool for the labeling and detection of enteric viruses (and their surrogates) which can be used to track virus behavior in situ.
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Rawsthorne et al. (2009) studied Enteric viruses. Fluorescent labeling strategy using streptavidin-labeled Quantum dots bound to biotinylated capsids vs. Control MNV-1 was evaluated on Relative fluorescence. A fluorescent labeling strategy using Quantum dots bound to biotinylated capsids yielded a relative fluorescence of 528 ± 48.1 for bio-MNV-1 compared to 112 ± 8.6 for control MNV-1.
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