A rapid detection method for Vaccinia virus, the surrogate for smallpox virus.
Prior to the World Health Organization's announcement of total eradication in 1977 [J. Am. Med. Assoc. 281 (1999) 1735], smallpox was a worldwide pathogen. Vaccinations were ceased in 1980 and now with a largely unprotected world population, smallpox is considered the ideal biowarfare agent [Antiviral Res. 57 (2002) 1]. Infection normally occurs after implantation of the virus on the oropharyngeal or respiratory mucosa [J. Am. Med. Assoc. 281 (1999) 2127]. Smallpox virus can be detected from the throats of exposed individuals prior to onset of illness and prior to the infectious stage of the illness. A rapid, sensitive real-time assay to detect Variola virus (smallpox) has been developed using the Vaccinia virus, a surrogate of smallpox, as a target. Cyanine 5 dye-labeled anti-Vaccinia antibody was used in a sandwich immunoassay to produce a fluorescent signal in the presence of the Vaccinia virus. The signal was detected using the Analyte 2000 biosensor (Research International, Monroe, WA). The Analyte 2000 uses a 635 nm laser diode to provide excitation light that is launched into a polystyrene optical waveguide. Fluorescent molecules within the evanescent wave are excited and a portion of their emission energy recouples into the waveguide. A photodiode quantifies the emission light at wavelengths between 670 and 710 nm. The biosensor was able to detect a minimum of 2.5 x 10(5) pfu/ml of Vaccinia virus in seeded throat culture swab specimens.
Department of Biology and Center for Biological Defense, University of South Florida, 4202 East Fowler Avenue, SCA 110, Tampa, FL 33620-5200, USA. kdonalds@cas.usf.edu
Kim A
KA
Donaldson
Marianne F
MF
Kramer
Daniel V
DV
Lim
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005336:Fiber Optic Technology / Q000295:instrumentation*; D016231:Fluorescence Polarization Immunoassay / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D009862:Online Systems; D010614:Pharynx / Q000821:virology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014616:Vaccinia virus / Q000302:isolation & purification*; D012901:Variola virus / Q000302:isolation & purification*