Abstract Background This paper outlines the methodology, study population and response rate of a third large Dutch population-based cross-sectional serosurvey carried-out in 2016/2017, primarily aiming to obtain insight into age-specific seroprevalence of vaccine-preventable diseases to evaluate the National Immunization Programme (NIP). In addition, Caribbean Netherlands (CN) was included, which enables additional research into tropical pathogens. Methods A two-stage cluster sampling technique was used to draw a sample of Dutch residents (0–89 years) (NS), including an oversampling of non-Western migrants, persons living in low vaccination coverage (LVC) areas, and an extra sample of persons born in Suriname, Aruba and the former Dutch Antilles (SAN). A separate sample was drawn for each Caribbean island. At the consultation hours, questionnaires, blood samples, oro- and nasopharyngeal swabs, faeces, − and only in the Netherlands (NL) saliva and a diary about contact patterns – were obtained from ...