The widespread use of gold nanoparticles (GNPs) as labels in diagnostics and detection is due to a unique combination of chemical and physical properties that allow biological molecules to be detected at low concentrations. In this critical review detection methods based on GNPs are divided up and discussed based on the way in which signals are generated in response to specific target molecules. Particular attention is devoted to methods that allow target molecules to be detected with the unaided eye because these, more than any other, harness the full range of properties that make GNPs unique. Methods that are discussed include those in which specific target molecules induce a visible colour change, chromatographic methods that allow non-specialized users to perform sophisticated tests without additional equipment and methods in which trace amounts of GNPs are rendered visible to the unaided eye by catalytic deposition of a metal such as silver. The use of metal deposition as a means of enhancing the signal for optical and electrical detection is also reviewed. The other detection methods included in this review are based on interactions between GNPs and molecules located in close proximity to their surface. These include methods in which light emission from such molecules is enhanced (surface enhanced Raman scattering) or quenched (fluorescence), and methods in which the accumulation of specific target molecules induce subtle changes in the extinction spectra of GNPs that can be followed in real time with inexpensive equipment (166 references).