Colorimetric lactate dehydrogenase (LDH) assay for evaluation of antiviral activity against bovine viral diarrhoea virus (BVDV) in vitro.
A rapid and sensitive screening assay has been established for in vitro evaluation of antiviral compounds against bovine viral diarrhoea virus (BVDV), which is widely used as a surrogate for hepatitis C virus (HCV). The procedure is based on photospectrometrical assessment for the viability of virus-infected cells via extracellular leakage of lactic dehydrogenase (LDH). The level of LDH in culture supernatants of BVDV-infected Madin-Darby bovine kidney (MDBK) cells was significantly higher than those of mock-infected MDBK cells. Under optimized assay conditions, the LDH level was found to correlate well with the degree of viral replication. When the 50% effective concentrations (EC50s) of ribavirin, cyclosporine A and human interferon-alpha for BVDV replication were determined by the established LDH method and compared with those obtained by a conventional tetrazolium colorimetric (MTT) method, there was a complete correlation in EC50s between the two methods. Furthermore, a much higher ratio of background activity (noise) to sample activity (signal) could be achieved with the LDH method than with the MTT method, indicating that the present LDH assay permits a sensitive, rapid and reliable screening of compounds for their anti-BVDV activity and may be useful for the discovery of novel anti-HCV agents.
Baba
Chiaki
C
Department of Dermatology, Graduate School of Medical & Dental Sciences, Kagoshima University, Kagoshima, Japan.
Yanagida
Koichiro
K
Kanzaki
Tamotsu
T
Baba
M
M
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002417:Cattle; D002478:Cells, Cultured; D003124:Colorimetry; D001908:Diarrhea Viruses, Bovine Viral / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000032:analysis*; D011994:Recombinant Proteins; D012254:Ribavirin / Q000494:pharmacology; D013778:Tetrazolium Salts; D013844:Thiazoles; D014779:Virus Replication / Q000187:drug effects / Q000502:physiology*
D000998:Antiviral Agents; D007370:Interferon Type I; D011994:Recombinant Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D012254:Ribavirin; D007770:L-Lactate Dehydrogenase; C022616:thiazolyl blue