Biosensors combine a transducer with a biorecognition element and thus are able to transform a biochemical event on the transducer surface directly into a measurable signal. By this they have the potential to provide rapid, real-time, and accurate results in a comparatively easy way, which makes them promising analytical devices. Since the first biosensor was introduced in 1962 as an "enzyme electrode" for monitoring glucose in blood, medical applications have been the main driving force for further biosensor development. In this chapter we outline potential biosensor setups, focusing on transduction principles, biorecognition layers, and biosensor test formats, with regard to potential applications. A summary of relevant aspects concerning biosensor integration in efficient analytical setups is included. We describe the latest applications of biosensors in diagnostic applications focusing on detection of molecular biomarkers in real samples. An overview of the current state and future trends of biosensors in this field is given.