Tetrazolium-based colorimetric assay for the detection of HIV replication inhibitors: revisited 20 years later.
Since its first description 20 years ago, the tetrazolium-based colorimetric (MTT) assay using MT-4 cells for the detection of anti-HIV compounds has been widely used. This method, which remains popular, provides more information than more recently developed methods and, therefore, represents a useful methodology on its own or in combination with other screening systems. The replication of HIV in MT-4 cells is usually monitored 5 d after infection; therefore, this protocol can be divided into three steps: the infection (at day 0), an incubation period (5 d) and the evaluation (at day 5). The long-standing and intensive use of the MTT method has taught users of the limitations and, equally importantly, the unexpected advantages of the MT-4/MTT assay. The use of this method can be extended to antiviral testing of compounds against other cyto-destructive viruses.
Pannecouque
Christophe
C
Rega Institute for Medical Research, Katholieke Universiteit Leuven, Minderbroedersstraat 10, B-3000 Leuven, Belgium. christophe.pannecouque@rega.kuleuven
Daelemans
Dirk
D
De Clercq
Erik
E
D019380:Anti-HIV Agents / Q000494:pharmacology*; D002461:Cell Line, Transformed; D003124:Colorimetry / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015497:HIV-1 / Q000187:drug effects; D015498:HIV-2 / Q000187:drug effects; D006801:Humans; D013778:Tetrazolium Salts
D019380:Anti-HIV Agents; D013778:Tetrazolium Salts; C042207:4-anisyltetrazolium blue