High throughput quantitative colorimetric microneutralization assay for the confirmation and differentiation of West Nile Virus and St. Louis encephalitis virus.
An automated colorimetric micro-neutralization assay (CmNt) was developed for confirmation and differentiation of West Nile Virus (WNV)-positive human sera as a higher throughput alternative to the standard six-well plaque-reduction neutralization test (PRNT). CmNt was performed in high-capacity 96-well micro-titer plates and required 4-6 days to complete. Inhibition of infection was determined by reduced neutral red-dye retention and conveniently recorded by a colorimetric plate reader. Human sera previously confirmed by PRNT as either negative (N = 52), WNV positive (N = 81), or St. Louis encephalitis virus positive (N = 12) were tested by CmNt; interpreted results were virtually identical to PRNT with a reduced turnaround time and higher throughput. Additionally, a handful of dengue virus positive and negative specimens (four each) were tested by CmNt; interpreted results were identical to PRNT.
Taketa-Graham
Michael
M
California Department of Public Health, Center for Infectious Disease, Richmond, CA, USA. mtaketa1@comcast.net
Powell Pereira
Jaime L
JL
Baylis
Elizabeth
E
Cossen
Cynthia
C
Oceguera
Leopoldo
L
Patiris
Peter
P
Chiles
Robert
R
Hanson
Carl V
CV
Forghani
Bagher
B
Forghani
BagHer
B
D000818:Animals; D002522:Chlorocebus aethiops; D003124:Colorimetry / Q000379:methods*; D004665:Encephalitis Virus, St. Louis / Q000302:isolation & purification*; D009500:Neutralization Tests / Q000379:methods*; D014709:Vero Cells; D014902:West Nile virus / Q000302:isolation & purification*