High-Throughput Characterization of Viral and Cellular Protein Expression Patterns During JC Polyomavirus Infection

Discussion The study of viral infections in vitro has provided innumerable advances to the field of virology. However, the lack of rapid and efficient screening tools has hindered research progress for some viruses, like JCPyV (Houff et al., 1983; Zu Rhein, 1983; Assetta and Atwood, 2017). To overcome this challenge, the development of high-throughput analyses is needed to help aid in the production of large data sets and generation of multiple lines of inquiry. Current methodologies for analyzing JCPyV infectivity predominantly rely on manual quantitation of infection by indirect immunodetection of viral proteins by epifluorescence microscopy. While these methodologies have provided crucial information into the characterization of JCPyV infectivity, the low-throughput nature of microscopy-based techniques has prevented the employment of large-scale screens and also hindered productivity within the field. Adapting the ICW procedure to effectively measure JCPyV infectivity has enabled faster analysis by improving efficiency by ∼9 person hours per 96-well plate in comparison to traditional FFU manual assessments. Moreover, this semi-automated assay allows for enhanced user objectivity. Utilization of ICW analysis to detect JCPyV infection and viral impacts on host-cell proteins provides a new technological platform for large-scale screens to measure the viral response to treatments such as inhibitors or siRNAs with enhanced efficiency and reduced use of resources. The ...
Source: Frontiers in Microbiology - Category: Microbiology Source Type: research