Abstract: The present invention provides microfluidic porous substrate-based devices for multiplexed biosensing. The devices are suitable for detecting viruses and bacteria, such as by way of detecting pathogenic genes and antibodies. The devices support reverse transcriptase loop-mediated isothermal amplification for rapid results within minutes. The devices also support vertical-lateral-vertical direction flow assays, such as in the form of a multi-layered adhesive bandage.