Biomolecular motors have been extensively studied as efficient molecular machines in detection systems owing to their unique signal conversion mechanisms and high energy conversion efficiencies. The application of these motors in the detection of pathogenic microorganisms is particularly promising. Through reasonable design and optimization, biomolecular motors can enable precise and efficient detection, enhancing clinical diagnostics. This paper reviews recent advances in detection systems utilizing various biomolecular motors, including kinesin, dynein, myosin, DNA polymerase, FoF1-ATPase, and flagellar motors. Detection mechanisms involving these motors are also introduced. Furthermore, the review covers recent progress in detecting antigens, antibodies, bacteria, and small molecules using biomolecular motors. Finally, the challenges and future prospects of biomolecular motor-based detection systems for pathogenic microorganisms are discussed, highlighting their potential as rapid and efficient tools for applications in food safety and medicine.