ATeam technology for detecting early signs of viral cytopathic effect

DOYSABAS, Karla Cristine C.; OBA, Mami; ISHIBASHI, Tomoki; SHIBATA, Hideki; TAKEMAE, Hitoshi; SHIMODA, Hiroshi; TARIGAN, Ronald; MIZUTANI, Tetsuya; IIDA, Atsuo; HONDO, Eiichi

Adenosine 5’-triphosphate (ATP), the major energy currency of the cell, is involved in many cellular processes, including the viral life cycle, and can be used as an indicator of early signs of cytopathic effect (CPE). In this study, we demonstrated that CPE can be analyzed using an FRET-based ATP probe named ATP indicator based on Epsilon subunit for Analytical Measurements (ATeam). The results revealed that as early as 3 hr, the virus infected cells showed a significantly different Venus/cyan fluorescent protein (CFP) ratio compared to the mock-infected cells. The ATeam technology is therefore useful to determine the early signs of ATP-based CPE as early as 3 hr without morphology-based CPE by light microscopy, and enables high throughput determination of the presence of microorganisms in neglected samples stored in laboratories.