Abstract: An integrated multiplexed acoustic wave biosensor chip system with enhanced sensitivity has been developed. The biosensor system incorporates one or more microfluidic channels, coated with target-specific binding films enabling rapid and early detection of viral, bacterial or parasitic targets such as Dengue virus and sexually transmitted diseases in specimens from potentially infected patients. The biosensors are used in portable analytical systems that are suitable for real-time point of care (POC) clinical diagnosis in cost sensitive and/or resource limited settings. The highly sensitive biosensors utilize thinned single crystal piezoelectric substrates that propagate layer guided shear horizontal acoustic plate mode (LG-SH-APM) waves in sensing regions bearing immobilized binders that provide simultaneous and direct detection of mass changes due to multiple bound target pathogens or molecules.