INTRODUCTION: Considering the usefulness of mobile health (mHealth) technologies in healthcare delivery in low- and middle-income countries, including Ghana; there is a need to explore healthcare professionals' perspectives on the availability and use of mHealth for disease screening and treatment of patients' conditions. The study's main aim is to explore healthcare professionals' perspectives regarding the availability and use of mHealth applications for disease screening and management at point-of-care in Ghana's Ashanti Region. MATERIALS AND METHODS: We conducted in-depth interviews with healthcare professionals who use mHealth applications daily between July and September 2020. A purposive sampling strategy was employed to select healthcare professionals who have been using mobile health application tools to support healthcare delivery. The researchers conducted 14 in-depth interviews with healthcare professionals on the availability and use of mHealth applications to support disease diagnosis and treatment of patients' conditions. Data were transcribed, coded, arranged, and analyzed to determine categories and themes. RESULTS: The study results demonstrated that healthcare workers had positive perceptions towards mHealth applications. Healthcare professionals identified significant challenges concerning mHealth applications: the high cost of data; lack of education or limited awareness; poor mobile networks; unstable internet connectivity; erratic power supply; and unavailability of logistics. Healthcare professionals identified the following prerequisite strategies to strengthen the use and scale-up of mHealth applications: stable internet connectivity; creating awareness; supplying logistics; reducing the cost of data; and developing local mobile apps. CONCLUSIONS: The study results revealed the availability of mHealth applications at the individual level for disease screening and treatment support of patients' conditions. The study also showed several significant challenges facing mHealth applications which need to be addressed to guarantee the successful implementation and scaling-up of mHealth activities at all levels of healthcare delivery. Hence, future research should incorporate healthcare professionals' perspectives to completely understand mHealth implementation and scaling-up challenges and measures to inform policy regulations.