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MICROPLATE READERS


VIRUS ASSAYS ON


Martin Mangold discusses improving effi ciency and decreasing costs


V


iruses, nowadays more than ever, pose growing threats to our health, society and economy. Detection systems


capable of analysing the mechanisms of viral infections as well as quantifying related diagnostic parameters are important tools to understand virus-host interactions and develop new treatments or diagnostic tests.


In the past, these applications


mainly revolved around microscopy- related techniques. Today more and more experiments are performed with microplate readers, as these off er several advantages in comparison to more traditional methods. On one hand, the microplate platform drastically reduces sample volume and accordingly assay cost. On the other, the higher number of tested samples per run reduces overall measurement times and enables a higher throughput. In addition, microplate readers provide a quantitative output that is less prone to interpretation or human error, compared to more qualitative, image-based readouts. Many microplate readers come also equipped with a


variety of detection modes, providing the possibility to measure a huge variety of diff erent assays on one device.


Accordingly, microplate readers have become an indispensable tool in virology research, covering a multitude of assays from basic to drug research. Roughly speaking, these assays can be categorised in three groups: functional virus assays, diagnostic assays, and anti-viral drug


24 www.scientistlive.com


MICROPLATE READERS


screening, which span the whole spectrum of detection modes from fl uorescence intensity to AlphaScreen.


FUNCTIONAL VIRUS ASSAYS Functional virus assays deal with the analysis of host-virus interactions. T ey are employed to clarify the cellular and molecular mechanisms behind infection and the role of the involved virus and host proteins. Generally, the infl uence of these interaction partners is investigated by gene knockout or inhibition and virus replication and viral gene expression are used as readouts. Furthermore, plate readers with atmospheric and temperature control enable real-time kinetic data acquisition of infection, replication and neutralisation assays monitored by fl uorescent or luminescent reporter proteins in living cells. Together with the measurement of cell viability, cytopathic eff ect (CPE) and TCID50, which can


The Clariostar Plus from BMG Labtech


easily be performed on a microplate


reader, this allows to study virus infectivity and replication directly in host cells.


DIAGNOSTIC ASSAYS Diagnostic assays are employed to effi ciently assess the presence of a virus and its spread in a population. For this purpose, assay sensitivity and speed are essential to allow for the fast and accurate evaluation of infection rates. T ese quantitative detection methods rely mainly on the detection of viral antigens or antibodies (ELISA) or of the viral genome (PCR, LAMP) in patient samples. One example is the loop-mediated isothermal amplifi cation (LAMP) assay which amplifi es viral gene material in a single 65°C incubation step (Fig. 1). LAMP products can be easily detected on a plate reader and the assay can be directly run and measured in real-time on plate readers with 65°C heating. While PCR and


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