Development of a versatile biosensing toolkit is in urgent need for rapid and multiplexed detection applications. In this work, an electronic particle counter-implemented versatile biosensing toolkit has been developed for detecting a range of targets with high sensitivity, broad detection range, multiplexibility, simple operation, and low cost. The electrical resistance-based particle counter conventionally measuring the number of microspheres (1-100 μm) can quantify analytes. The versatility of this approach is verified by assaying small molecules, protein biomarkers, pathogen bacteria, and tumor cells using three strategies: (1) antigen-antibody interaction, (2) DNA hybridization, and (3) polypeptide recognition. More importantly, this biosensing toolkit allows the simultaneous detection of multiple targets with a broad detection range from pg mL-1 to μg mL-1, showing great potential as a powerful technique for food safety testing and biomedical diagnosis.