A Semi-automated High-Throughput Microtitration Assay for Filoviruses

Claire Marie Filone; David Miller; Victoria Wahl-Jensen

doi:10.1007/978-1-4939-7116-9_12

A Semi-automated High-Throughput Microtitration Assay for Filoviruses

Methods Mol Biol. 2017:1628:163-175. doi: 10.1007/978-1-4939-7116-9_12.

Authors

Claire Marie Filone¹, David Miller¹, Victoria Wahl-Jensen²

Affiliations

¹ Department of Homeland Security, National Biological Threat Characterization Center, National Biodefense Analysis & Countermeasures Center, Science & Technology Directorate, Fort Detrick, MD, USA.
² Department of Homeland Security, National Biological Threat Characterization Center, National Biodefense Analysis & Countermeasures Center, Science & Technology Directorate, Fort Detrick, MD, USA. victoria.jensen@nbacc.dhs.gov.

PMID: 28573618
DOI: 10.1007/978-1-4939-7116-9_12

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.

Keywords: Assay; Biosafety level 4 (BSL-4); Dilution; Ebola; Efficiency; Endpoint; Filovirus; High-throughput screening (HTS); Liquid handling; Luminescence; Luminescent; Marburg; Microtitration; Semi-automated; Tissue culture infectious dose (TCID50); Viability.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Survival / genetics
Filoviridae / genetics
Filoviridae / pathogenicity*
High-Throughput Screening Assays / methods*
Humans