Abstract
The 50% tissue culture infectious dose (TCID50) endpoint dilution assay is one of the gold standard methods for measuring filovirus infectivity. We have increased virology microtitration assay throughput at biosafety level (BSL)-4 by implementing automated liquid handling and semi-automated assay endpoint readout. Utilization of automated liquid handling for cell plating and virus dilution along with optimization of the assay endpoint readout, using a luminescent-based cell viability assay and an automated plate reader, has improved workflow efficiency, reduced operator burden and assay time, decreased assay variability, and increased data return.
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Acknowledgment
This work was funded under Contract No. HSHQDC-07-C-00020 awarded by the Department of Homeland Security (DHS) Science and Technology Directorate (S&T) for the management and operation of the National Biodefense Analysis and Countermeasures Center (NBACC), a Federally Funded Research and Development Center. The views and conclusions contained in this document are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the DHS or S&T. In no event shall DHS, NBACC, S&T, or Battelle National Biodefense Institute have any responsibility or liability for any use, misuse, inability to use, or reliance upon the information contained herein. DHS does not endorse any products or commercial services mentioned in this publication.
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Filone, C.M., Miller, D., Wahl-Jensen, V. (2017). A Semi-automated High-Throughput Microtitration Assay for Filoviruses. In: Hoenen, T., Groseth, A. (eds) Ebolaviruses. Methods in Molecular Biology, vol 1628. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7116-9_12
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DOI: https://doi.org/10.1007/978-1-4939-7116-9_12
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