Optimization of the interferon assay using inhibition of Semliki forest virus-ribonucleic acid synthesis.
The method for assaying chicken interferon by its inhibition of viral ribonucleic acid (RNA) synthesis was optimized for the chicken embryo fibroblast-Semliki Forest virus (SFV) system, with respect to time, multiplicity of infection, and addition of actinomycin D and (3)H-uridine. Incorporation of (3)H into viral RNA is reproducible, and amounts to 10(4) to 2 x 10(4) counts per min per uninhibited culture per 1 muCi per 6 hr. The assay may be carried out in less than 1 day and is sensitive (0.05 units/ml) and exact (+/-20% of the mean titers on different days); it can be used for purification procedures as well.
D000818:Animals; D002642:Chick Embryo; D002850:Chromatography, Gel; D046508:Culture Techniques; D003609:Dactinomycin / Q000494:pharmacology; D005347:Fibroblasts / Q000382:microbiology; D007372:Interferons / Q000032:analysis* / Q000494:pharmacology; D008722:Methods; D008851:Micropore Filters; D009522:Newcastle disease virus; D012367:RNA, Viral / Q000096:biosynthesis*; D012672:Semliki forest virus / Q000254:growth & development / Q000378:metabolism*; D013997:Time Factors; D014316:Tritium; D014529:Uridine / Q000378:metabolism; D010948:Viral Plaque Assay
D012367:RNA, Viral; D014316:Tritium; D003609:Dactinomycin; D007372:Interferons; D014529:Uridine