A fully automatic approach to locate polyomavirus particles in transmission electron microscopy images is presented that can localize intact particles, many damaged capsids, and an acceptable percentage of superposed ones. Performance of the approach is quantified in 25 electron micrographs containing nearly 390 particles and compared with the interpretation of the micrographs by two independent electron microscopy experts. All parameterization is based on the particle expected dimensions. This approach uses indicators calculated from the local co-occurrence matrix of gray levels to assess the textured pattern typical of polyomavirus and prune the initial set of candidates. In more complicated backgrounds, about 2–10% of the elements survive. A restricted set of the accepted points is used to evaluate the typical average and variance and to reduce the set of survivors accordingly. These intermediate points are evaluated using (i) a statistical index concerning the radiometric distribution of a circular neighborhood around the centroid of each candidate and (ii) a structural index resuming the expected morphological characteristics of eight radial intensity profiles encompassing the area of the possible particle. This hierarchical approach attains 90% efficiency in the detection of entire virus particles, tolerating a certain lack of differentiation in the borders and a certain amount of shape alterations.