ArticleParasites & vectors2015
Development of a Real-Time Cell Analysing (RTCA) method as a fast and accurate screen for the selection of chikungunya virus replication inhibitors.
Article in Parasites & vectors, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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12 citing papers in PubMed.
- Beyond Tuberculosis: Unlocking the Antifungal Potential of Novel Isoniazid Derivatives against Non-ACS omega · 2026Article
- Evaluation of three alternative methods to the plaque reduction neutralizing assay for measuring neutralizing antibodies to dengue virus serotype 2.Virology journal · 2024Article
- Dual effect of aucubin on promoting VEGFR2 mediated angiogenesis and reducing RANKL-induced bone resorption.Chinese medicine · 2023Article
- DNA-encoded chemical libraries yield non-covalent and non-peptidic SARS-CoV-2 main protease inhibitors.Communications chemistry · 2023Article
- Prophylactic strategies to control chikungunya virus infection.Virus genes · 2021Review
- Application of xCELLigence real-time cell analysis to the microplate assay for pertussis toxin induced clustering in CHO cells.PloS one · 2021Article
- Real-time cell analysis system in cytotoxicity applications: Usefulness and comparison with tetrazolium salt assays.Toxicology reports · 2020Review
- Structure-based design generated novel hydroxamic acid based preferential HDAC6 lead inhibitor with on-target cytotoxic activity against primary choroid plexus carcinoma.Journal of enzyme inhibition and medicinal chemistry · 2019Article
- Article
- Exploring the Potential of Electrical Impedance Tomography for Tissue Engineering Applications.Materials (Basel, Switzerland) · 2018Article
- Development of a Real-Time Cell Analysis (RTCA) Method as a Fast and Accurate Method for Detecting Infectious Particles of the Adapted Strain of Hepatitis A Virus.Frontiers in cellular and infection microbiology · 2018Article
- Advanced Glycation End Products Inhibit the Proliferation of Human Umbilical Vein Endothelial Cells by Inhibiting Cathepsin D.International journal of molecular sciences · 2017Article
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4 authors.
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Abstract
backgroundThe xCELLigence real-time cell analysis (RTCA) system is an established electronic cell sensor array. This system uses microelectronic biosensor technology that is verified for real-time, label-free, dynamic and non-offensive monitoring of cellular features, including detection of viral cytopathic effect (CPE). Screening viral replication inhibitors based on presence of CPE has been applied for different viruses, including chikungunya virus (CHIKV). However, most CPE-based methods, including MTT and MTS assays, do not provide information on the initiation of CPE nor the changes in reaction rate of the virus propagation over time. Therefore, in this study we developed an RTCA method as an accurate and time-based screen for antiviral compounds against CHIKV.
methodsCHIKV-infected Vero cells were used as an in vitro model to establish the suitability of the RTCA system as a quantitative analysis method based on the induction of CPE. We also performed an MTS assay as a CPE-based conventional method. Experimental assays were carried out to evaluate the optimal seeding density of the Vero cells, cytotoxicity of the tested compounds, titration of CHIKV and the antiviral activity of ribavirin, which has been reported as an effective compound against CHIKV in vitro replication.
resultsThe optimal time point for viral inoculation was 18 h after seeding the cells. We determined that the maximum non-toxic dose (MNTD) of ribavirin was 200 μg/ml for Vero cells. Regarding the dynamic monitoring of Vero cell properties during antiviral assay, approximately 34 h post-infection, the normalised Cell Index (CI) values of CHIKV-infected Vero cells started to decrease, while the vehicle controls did not show any significant changes. We also successfully showed the dose dependent manner of ribavirin as an approved in vitro inhibitor for CHIKV through our RTCA experiment.
conclusionRTCA technology could become the prevailing tool in antiviral research due to its accurate output and the opportunity to carry out quality control and technical optimisation.
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