| Abstract
| - Virus detection and enumeration has become increasinglyimportant in fields ranging from medicine and biotechnology to environmental science. Although there are awide variety of techniques that can be used to countviruses, there is demand for a rapid and more accuratemeans for virus enumeration. In this work, the performance of a flow cytometer that was designed and custom-built specifically for rapid detection of single viruses wasevaluated. The instrument, designated a single nanometricparticle enumerator (SNaPE), was characterized andcalibrated using fluorescent polystyrene nanospheres. Thereliability of the instrument with respect to virus enumeration was demonstrated for three medically relevantviruses, adenovirus-5, respiratory syncytial virus, andinfluenza A, treated with a fluorescent nucleotide stain.In each case, the SNaPE yielded a virus particle concentration consistent with, but slightly lower than, transmission electron microscopy (TEM) results, as expected. Inaddition, on the basis of calibration of signal intensity, theaverage peak height for a given virus was correlated withgenome size, as expected. In contrast to time-consuminganalyses such as TEM and plaque titers, SNaPE analysisof pure virus samples (including sample handling, datacollection, and data processing) can be completed within1 h.
|
