Studies into the systematics of wolf spiders have mainly employed morphological characters of adult spiders, in particular features of the male and female genitalia, and more recently mitochondrial DNA sequence data. However, there is still no established phylogenetic framework for the Lycosidae, even at the subfamily level. This study uses a novel morphological character set, the chaetotaxy of lycosid larvae (presence and arrangement of setae and slit organs), to infer systematic information on seven species of wolf spiders that are currently listed in three subfamilies: Lycosinae [Alopecosa pulverulenta (Clerck 1757), Hogna antelucana (Montgomery 1904), Rabidosa rabida (Walckenaer 1837), Trochosa ruricola (DeGeer 1778)], Piratinae [Hygrolycosa rubrofasciata (Ohlert 1865), Pirata hygrophilus (Clerck 1757)], and Sosippinae (Sosippus californicus Simon 1898). Cheliceral and tarsal (legs I and II) chaetotaxic patterns of the first postembryo showed equivalent chaetotaxic complexes amongst all species but revealed considerable differences between representatives of the three subfamilies. Sosippus californicus showed the most complex pattern and P. piraticus the most reduced arrangement. In addition, it casts doubt on the previous listings of H. rubrofasciata in either the Lycosinae or Piratinae, as its chaetotaxic setae arrangement was more similar to S. californicus than to any other species investigated here.