Multiple-Channel Funnel Adapted Sensing Tube (MFAST) for the Simple and Duplex Detection of Parasites

Abstract

Leishmaniasis poses a significant global health threat, infecting millions of people annually, particularly in tropical and subtropical regions. Timely and accurate detection of the Leishmania species is crucial for effective treatment and control of this debilitating disease. This study introduces the multi-channel funnel adapted sensing tube (MFAST) chip, a simple diagnostic tool designed for the rapid detection of Leishmania panamensis. MFAST is fabricated through 3D printing and sacrificial molding of acrylonitrile butadiene styrene (ABS) and the reagents are transported between reservoirs by gravity. We combine experiments and finite element analysis to facilitate smoother fluid flow, improving the overall performance of the device. Highly sensitive and specific RPA-CRISPR/Cas12a assay is utilized in the chip, achieving a detection limit as low as 1000 parasites/mL (detecting as few as 5 parasites per reaction). The multichannel design enables duplex detection, allowing for simultaneous identification of both L. braziliensis and L. panamensis through distinct channels. Furthermore, stability tests indicate that lyophilized reagents retain functionality for up to 15 days when stored at 4 °C, underscoring the potential of this chip for practical diagnostic applications in low-resource settings.