Nanotechnology-based paper microfluidics for rapid point-of-care detection and differentiation of snake venom types

Snake envenomation is recognized as a neglected tropical disease, contributing to high mortality rates and causing significant organ damage, particularly to the liver, kidneys, and brain. The primary treatment involves administering antivenom, which consists of polyclonal antibodies developed against various snake venoms. However, antivenom therapy can lead to serum-related complications, reducing its effectiveness. Therefore, targeting specific therapeutic molecules could significantly improve snake envenomation treatment. Identifying the snake species is a major challenge due to their similar morphological characteristics. Globally, only two snake venom diagnostic kits are available that have been developed to detect country-specific snake venom. Hence, there is an urgent need to develop new diagnostic assays tailored for detecting venom specific to India. To address this, the current study focuses on detecting functional enzyme components of venomous snake species, such as phospholipase A2, hyaluronidase, and proteases. The study is based on the loading of the dye-loaded stimuli-responsive nanoparticles, including liposomes (sensitive to phospholipase A2), hyaluronic acid-chitosan nanoparticles (sensitive to hyaluronidase), and casein nanoparticles (sensitive to proteases) into paper-based microfluidics and tested with various snake venoms. The device successfully detects and distinguishes between wet bites and dry bites, as well as viper and elapid species. • Developed nanoparticle-based paper microfluidics for the differentiation of viper and elapid snake envenomation. • Optimization and characterisation of stimuli-responsive nanoparticles. • The sensing of the venom is based on the dye release from the stimuli-responsive nanoparticles and enzymes in the venom. • Biosensors exhibit high sensitivity and selectivity to the viper and elapid venom. • The device is discarded safely by incineration after sensing.