Tracking Ebolavirus genomic drift in a replicating virus with a resequencing microarray, verification with NGS 2

This study demonstrates the ability of an Ebolavirus resequencing microarray to determine the sequence of the Zaire ebolavirus glycoprotein that has been engineered into the place of the surface protein of a recombinant vesicular stomatitis virus expressing green fluorescent protein (rVSV-EBOVgp-GFP). The rVSV-EBOVgp-GFP was cultured in VERO-E6 cells for three passages either in the presence of a monoclonal antibody that blocks infection (KZ52) or in control cultures with no antibody. Culture supernatant and cell lysate was collected before passaging and after each passage. RNA was extracted from each sample and the sequence of the Ebola glycoprotein in each sample was determined by the Ebola resequencing microarray. Illumina Next Generation Sequencing was performed on the initial virus stock, before passaging and on the third passage of the KZ52 antibody selected virus stock to validate the microarray sequence results.