Development of an Automated Viral Plaque Assay Program and ELISpot Counting Program on 96-Well Plate Platform

Abstract

Plaque assay is a unique method and specialized measurement by virologists that provides a drag discovery until vaccine development. Also, the ELISpot assay is one of the processes that doctors use to detect CTLs to diagnose the patient. The approach of plaque assays and ELISpot assays is challenging and complicated. Both need experts and time to analyze by physical counting in well-plates, which causes abstract and stagnant to the expert. Hence, to dimmish the load of the specialists to physical counting in well plates, we have developed an automated platform to capture images from 96-well plates and do automate assay for both methods. The platform contains two major parts: Hardware used for acquiring images by a modification of IAI Tabletop with USB microscope and light souses and Software for image viral counting, that is simple to use and easy to setup. The software used for automated assays consists of two parts: the ELISpot assay part utilizes the theory of dynamic thresholding algorithms. The plaque assay uses unsupervised learning algorithms named k-mean clustering. The graphic user interface displays images of a virtual plaque and ELISpot, the virtual plaque assays, and the ELISpot assay results. The ELISpot counting algorithm was evaluated with the golden standard with 80% accuracy. The average correction of Plaque assay measurements by the machine is 94%.