A Modified Sample Preparation Protocol for High-Efficiency Lab-on-a-Disk-Based Separation and Single-Image Quantification of Soil-Transmitted Helminth Parasite Eggs in Stool.

Soil-transmitted helminths (STHs) present a significant global health challenge, particularly in tropical and subtropical regions. The current diagnostic standard involves the microscopic examination of a stool smear but it lacks sensitivity to detect infections of low intensity. Innovative solutions like lab-on-a-disk (LoD) technologies are emerging, showing promise in detecting low-intensity infections. Field tests conducted using our SIMPAQ (single-image parasite quantification) LoD device have demonstrated its potential as a diagnostic tool, especially for such low-intensity infections. Nevertheless, the device's efficiency has been limited by significant egg loss during sample preparation, low capture efficiency of eggs within the Field of View (FOV), and the presence of larger fecal debris that obstructs effective egg trapping and imaging. In this study, we conducted a set of laboratory experiments using model polystyrene particles and purified STH eggs to improve the sample preparation protocol. These experiments include the entire SIMPAQ procedure starting from sample preparation, infusing it into the LoD device, centrifugation, delivering the (model) eggs to the FOV, capturing an image, and analyzing it. We analyzed egg losses at each step of the procedure following the "standard" protocol, then elaborated and tested alternative, more efficient procedures. The resulting modified protocol significantly minimized particle and egg loss and reduced the amount of debris in the disk, thus enabling effective egg capture and clear images in the FOV, increasing the reliability of the diagnostic results.