Given the causal involvement of high-risk human papillomaviruses (HPVs) in cervical cancer and a subset of squamous cell carcinomas of other anogenital regions as well as the oropharynx, much attention has been focused on the development and application of HPV detection assays. HPV detection assays are almost exclusively based on the detection of viral nucleic acids, mostly viral DNA. The HPV detection methods that are nowadays in use can broadly be subdivided into target amplification methods and signal amplification methods. In this review, several principles of various methodologies are explained and examples of some commonly used HPV detection assays are given. In addition, attention is paid to the use of HPV assays for detecting clinically meaningful HPV infections, i.e. infections related to (pre)cancerous lesions, e.g. cervical cancer screening purposes. For the latter, it is important that HPV tests are clinically validated according to validation strategies as outlined in guidelines.