title: A colorimetric assay for viral agents that produce cytopathic effects.
abstract: Many animal viruses produce cytopathic effects in their host cells during a productive infection. While some virus infections can be assayed by the production of plaques, many viruses, while producing cytotoxicity, do not easily form plaques, or do not form plaques at all. Additionally, viruses within families such as the parvoviruses may have different preferred forms of titration making comparative virology difficult even among related groups. Porcine parvovirus (PPV), canine parvovirus (CPV), and minute virus of mice (MVM) are usually titrated using different infectivity assays. A direct comparison of infectious virus titer between these parvoviruses was sought, and a tetrazolium salt assay, MTT has been applied to measure cytopathic effect produced by viral infection for different members of the parvovirus family. Infectious PPV measured using the MTT and the TCID50 assays exhibited excellent correlation and titers for CPV and MVM were consistently duplicated using the MTT assay. The MTT assay was also applied to an unrelated virus, Sindbis, which is routinely titrated by plaque assay. MTT titration of Sindbis virus mutants was found to be valuable for preliminary screening. This assay can be adapted, by correlation to an accepted titration method, to any viral system which produces measurable cytopathic effect.
authors.affiliation: Department of Chemical and Biomolecular Engineering, North Carolina State University, 911 Partners Way, Raleigh NC 27695-7905, USA.
authors.forename: Caryn L
authors.initials: CL
authors.lastname: Heldt
authors.forename: Raquel
authors.initials: R
authors.lastname: Hernandez
authors.forename: Usharani
authors.initials: U
authors.lastname: Mudiganti
authors.forename: Patrick V
authors.initials: PV
authors.lastname: Gurgel
authors.forename: Dennis T
authors.initials: DT
authors.lastname: Brown
authors.forename: Ruben G
authors.initials: RG
authors.lastname: Carbonell
mesh_terms: D000818:Animals; D002460:Cell Line; D003124:Colorimetry / Q000379:methods*; D006224:Cricetinae; D003588:Cytopathogenic Effect, Viral; D004285:Dogs; D005562:Formazans / Q000378:metabolism; D006801:Humans; D008915:Minute Virus of Mice / Q000472:pathogenicity; D017993:Parvovirus, Canine / Q000472:pathogenicity; D028323:Parvovirus, Porcine / Q000472:pathogenicity; D015203:Reproducibility of Results; D012845:Sindbis Virus / Q000472:pathogenicity; D013552:Swine; D013778:Tetrazolium Salts / Q000378:metabolism; D010948:Viral Plaque Assay; D014780:Viruses / Q000472:pathogenicity*
chemical_list: D005562:Formazans; D013778:Tetrazolium Salts; C079782:MTT formazan