Portable applications of fluorescence detection systems have gained much attention in various fields and require system components to be small and compact. In this work, we report on a compact fluorescence detection system and demonstrate its application for fluorescence sensing and imaging. The light source and filter are integrated on a single chip for the proposed system, which not only realizes the separation between excitation and fluorescent lights but also improves the light-emitting diode (LED) light extraction efficiency. Furthermore, the detection system allows for a removable sample unit. The results indicate that the performance of the distributed Bragg reflector (DBR) filter based on an amorphous dielectric film is excellent with selection ratios larger than 4600:1. The peak emission wavelength of the LED is 528 nm. The influence of green light leakage can be neglected, and the fluorescent red light is dominant when the fluorescence detection system is used for sensing and imaging. The low-cost and monolithic DBR-integrated III-nitride LED chip makes the proposed architecture a competitive candidate for portable fluorescence detection applications.