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Updated: Mar 7, 2026
Virus Propagation and Cell-Based Colorimetric Quantification
Published on: April 7, 2023
Development of a high-throughput colorimetric Zika virus infection assay
Janis A Müller1, Mirja Harms1, Axel Schubert2
1Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.
Abstract:
Zika virus (ZIKV) is an emerging pathogen that causes congenital infections which may result in birth defects, such as microcephaly. Currently, no approved treatment or vaccination is available. ZIKV can be readily detected in cell culture where virally infected cells are normally stained by specific antibodies. As ZIKV regularly causes a cytopathic effect, we were wondering whether this viral property can be used to quantitatively determine viral infectivity. We here describe the use of an 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide-(MTT)-based cell viability assay that allows to determine ZIKV-induced cell death. We show that this colorimetric assay quantifies ZIKV infection over a broad range of viral dilutions in both monkey and human cells. It allows to determine inhibitory activities of antivirals that block ZIKV or to define the neutralizing antibody titers of ZIKV antisera. This MTT-based ZIKV detection assay can be evaluated by naked eye or computational tools, has a broad linear range, does not require large equipment or costly reagents, and thus represents a promising alternative to antibody-based assays, in particular in resource-poor settings. We propose to use this simple, fast, and cheap method for quantification of ZIKV neutralizing antibodies and testing of antiviral compounds.
Insights
A new MTT-based assay quantifies Zika virus (ZIKV) infection by measuring cell death. This simple, cost-effective method aids in testing antiviral drugs and ZIKV neutralizing antibodies, especially in resource-limited settings.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Zika virus (ZIKV) is an emerging pathogen linked to severe congenital abnormalities like microcephaly.
- Current therapeutic and preventative options for ZIKV infection are limited.
- Conventional ZIKV detection relies on antibody staining, which can be resource-intensive.
Purpose of the Study:
- To develop and validate a quantitative method for assessing ZIKV infectivity using cell viability.
- To establish a cost-effective and accessible assay for ZIKV detection and characterization.
Main Methods:
- Utilized a 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide (MTT)-based colorimetric assay to measure ZIKV-induced cell death.
- Evaluated the assay's performance in quantifying ZIKV infection across various viral dilutions in both monkey and human cell lines.
- Assessed the assay's utility in determining the efficacy of antiviral compounds and the titers of ZIKV-neutralizing antibodies.
Main Results:
- The MTT-based assay accurately quantifies ZIKV infection and cell death over a wide range of viral dilutions.
- The assay demonstrated effectiveness in both monkey and human cell lines.
- Demonstrated the assay's capability to determine antiviral activity and neutralizing antibody titers.
Conclusions:
- The MTT-based cell viability assay provides a simple, rapid, and inexpensive method for quantifying ZIKV infectivity.
- This assay is a promising alternative to traditional antibody-based methods, particularly for resource-poor settings.
- The developed assay facilitates the screening of antiviral compounds and the measurement of ZIKV-neutralizing antibodies.
