Improvement in Throughput and Accuracy ofNeutralization Assay Results by the Automation ofImage Acquisition and Analysis

Quantification neutralizing antibodies methods are crucial in the development and evaluation of new antiviral vaccines effectiveness and the assay most commonly used is the Plaque-Reduction Neutralization Test (PRNT). However, PRNT and its variations like FRNT (or microPRN) are time-consuming, not amenable to a large number of samples and difficult to adapt to a high throughput routine. Furthermore, the assay tends to have large variations among different analysts due to visual plaque recognition and manual counting errors. Therefore, providing a faster and more accurate counting for the neutralizing antibodies assays would minimize human inherent variations. Thus, innovations such as the automation at critical steps can increase throughput, reduce rework and generate more accurate results for vaccines developing and diagnosis. Companies specialized in microscopy, robotics and information technology developed robust systems able to standardize the image acquisition and processing for neutralization assays, which could aid the public health response to emerging viral diseases. In this context, automated systems have been used to obtain images and to identify morphological patterns setting the best ranges for the structures of interest, such as circularity, size, color intensity and other attributes.