Abstract
We present a lab-on-a-disk technology for fast identification and quantification of parasite eggs in stool. We introduce a separation and packing method of eggs contained in 1 g of stool, allowing for removal of commonly present solid particles, fat droplets and air bubbles. The separation is based on a combined gravitational and centrifugal flotation, with the eggs guided to a packed monolayer, enabling quantitation and identification of subtypes of the eggs present in a single field of view (FOV). The prototype was tested with stool samples from pigs and humans infected with intestinal parasites (soil-transmitted helminths eggs). The quality of the images created by this platform was appropriate for identification and quantification of egg types present in the sample.
Highlights
According to World Health Organization (WHO), more than 1.5 billion people, or 24% of the world’s population, are infected with soil-transmitted helminths (STH) worldwide [1]
The separation is based on a combined gravitational and centrifugal flotation, with the eggs guided to a packed monolayer, enabling quantitation and identification of subtypes of the eggs present in a single field of view (FOV)
The prototype was tested with stool samples from pigs and humans infected with intestinal parasites
Summary
According to World Health Organization (WHO), more than 1.5 billion people, or 24% of the world’s population, are infected with soil-transmitted helminths (STH) worldwide [1]. The Cornell Wisconsin method has been compared to several flotation-based devices that were developed so far and was systematically categorized as being less sensitive than its successor technologies, namely McMaster, FLOTAC and FECPAK as the most successful implementations [11]. The main limitations of the FLOTAC technique are the complexity of the application and the requirement for centrifugation of the sample using a large swinging bucket centrifuge, which is often not available in laboratories in developing countries. In FECPAK, a special tube with a central pillar is filled with a stool sample dissolved in flotation solution, allowing the parasite eggs to accumulate into a single viewing area within a fluid meniscus. A tube with a central pillar is filled with a stool sample dissolved in flotation solution, allowing the parasite eggs to accumulate into a single viewing area within a fluid meniscus. Centrifugal microfluidics is mostly used in, e.g., disc-based PCR [19,20], quantifying original levels of genomic material [21], plasma separation [22,23], ELISA tests [24,25,26], colorimetric assays for the determination of alcohol or glucose concentration in whole blood [27,28] culturing roundworms [29,30], water, food and soil analysis [31,32,33], whole blood cells isolation [34,35] and in many other applications reviewed by, e.g., Gorkin et al [36] and Strohmeier et al [37]
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