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Tetrazolium-based colorimetric assay for the detection of HIV replication inhibitors: revisited 20 years later

Nature Protocols volume 3, pages 427–434 (2008)Cite this article

Abstract

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.

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Figure 1: Preliminary assessment of the cytotoxicity of DMSO and NMP in MT-4 cells using the MTT/MT-4 assay.
Figure 2: Influence of the amount of viral inoculum on the production of the HIV stock.
Figure 3: Typical examples of results obtained with antiretroviral drugs nevirapine, an NNRTI; AZT, an NRTI and ritonavir, a protease inhibitor.

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Acknowledgements

We thank Mrs Kristien Erven for critical reading and fine editorial assistance.

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  1. Rega Institute for Medical Research, Katholieke Universiteit Leuven, Minderbroedersstraat 10, Leuven, B-3000, Belgium

    Christophe Pannecouque, Dirk Daelemans & Erik De Clercq

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  1. Christophe Pannecouque
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Correspondence to Christophe Pannecouque.

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Pannecouque, C., Daelemans, D. & De Clercq, E. Tetrazolium-based colorimetric assay for the detection of HIV replication inhibitors: revisited 20 years later. Nat Protoc 3, 427–434 (2008). https://doi.org/10.1038/nprot.2007.517

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