A novel and rapid method to quantify cytolytic replication of picornaviruses in cell culture.
Determining viral titers is a key issue in a wide variety of studies regarding different aspects of virology. The standard methods used for determining picornavirus titers are endpoint titration assay and plaque assay, both time consuming and laborious. The method described uses the tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide) that is reduced to formazane by cellular dehydrogenase, genes shown to be down-regulated during picornavirus infection. The amount formazane produced correlates with the viral titers obtained and can easily be measured using an ELISA plate reader. The colorimetric method has been evaluated using virus types from different genera of the Picornaviridae family. The MTT method reduces the time spent on determining the viral titers and still maintains a reliable accuracy.
Andersson
Per
P
Department of Chemistry and Biomedical Sciences, University of Kalmar, Smalandsgatan 24, S-39182 Kalmar, Sweden.
Alm
Stina
S
Edman
Kjell
K
Lindberg
A Michael
AM
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D003124:Colorimetry; D003588:Cytopathogenic Effect, Viral; D005562:Formazans / Q000378:metabolism; D006801:Humans; D010088:Oxidoreductases / Q000032:analysis / Q000378:metabolism; D010849:Picornaviridae / Q000302:isolation & purification* / Q000502:physiology; D012680:Sensitivity and Specificity; D013778:Tetrazolium Salts / Q000378:metabolism; D014773:Virology / Q000379:methods*; D014779:Virus Replication
D005562:Formazans; D013778:Tetrazolium Salts; D010088:Oxidoreductases