Simian virus 40 (SV40) is known to cause tumourigenesis. The main types of tumour induced by SV40 in laboratory animals mirror the human cancers that have been found to contain SV40 DNA or the viral oncoprotein. Increasing amounts of data support the notion that SV40 may be an aetiological factor in the development of human cancers. Retrospective birth cohort studies have been used in attempts to refute the alleged causal link between SV40 and human cancers. However, these observational studies are affected by several important confounding factors, which mean that firm conclusions cannot be drawn. In this essay, we consider the unique features of SV40 infection in humans and examine the limitations of conventional studies that seek to disprove the aetiological link with human cancer.