Digital bioanalysis places great emphasis on the highly sensitive and rapid detection of biomolecules at the single-molecule level. Rooted in single-molecule biophysics, this innovative approach offers numerous insights into biomolecular mechanisms with an unprecedented level of sensitivity and precision. Moreover, this method has significant potential to contribute to disease diagnostics, enabling the highly sensitive detection of biomarkers or pathogens for early disease diagnosis and continuous disease monitoring. However, the notable cost of detection and specialized equipment required for fabricating microdevices pose a challenge to accessibility and ease of use. This lack of versatility hinders the widespread adoption of digital bioanalysis. Here, we aim to illuminate the essential requirements for versatile digital bioanalysis and present prospects for biomedical applications that can be facilitated by attaining such versatility.