Development of a high-throughput colorimetric Zika virus infection assay.
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.
Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.
Janis A
JA
Müller
Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.
Mirja
M
Harms
Institute of Virology, Ulm University Medical Center, Ulm, Germany.
Axel
A
Schubert
Institute of Epidemiology and Medical Biometry, Ulm University, Ulm, Germany.
Benjamin
B
Mayer
Bernhard Nocht Institute for Tropical Medicine, World Health Organization Collaborating Centre for Arbovirus and Hemorrhagic Fever Reference and Research, Hamburg, Germany.|German Centre for Infection Research, Partner sites Hamburg-Luebeck-Borstel, Hamburg, Germany.
Stephanie
S
Jansen
CBMIT CNRS UMR-8601, Université Paris Descartes, CICB, Paris, France.
Jean-Philippe
JP
Herbeuval
Institute of Virology, Ulm University Medical Center, Ulm, Germany.
Detlef
D
Michel
Institute of Virology, Ulm University Medical Center, Ulm, Germany.
Thomas
T
Mertens
Departments of Virology and Immunology, and Veterinary Biosciences, University of Helsinki and Helsinki University Hospital, Helsinki, Finland.
Olli
O
Vapalahti
Bernhard Nocht Institute for Tropical Medicine, World Health Organization Collaborating Centre for Arbovirus and Hemorrhagic Fever Reference and Research, Hamburg, Germany.|German Centre for Infection Research, Partner sites Hamburg-Luebeck-Borstel, Hamburg, Germany.
Jonas
J
Schmidt-Chanasit
Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany. jan.muench@uni-ulm.de.
Jan
J
Münch
D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D002460:Cell Line; D002470:Cell Survival; D002522:Chlorocebus aethiops; D003124:Colorimetry / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D016898:Interferon-alpha / Q000494:pharmacology; D009500:Neutralization Tests / Q000379:methods; D014709:Vero Cells; D000071244:Zika Virus / Q000187:drug effects / Q000276:immunology; D000071243:Zika Virus Infection / Q000175:diagnosis* / Q000276:immunology / Q000821:virology*
D057134:Antibodies, Neutralizing; D007106:Immune Sera; D016898:Interferon-alpha
MTT; PRNT; Screening; ZIKV; Zika virus