In this study, 200 Escherichia coli isolates from 22 rainwater tank samples in Southeast Queensland, Australia, were tested for the presence of 20 virulence genes (VGs) associated with intestinal and extraintestinal pathotypes. In addition, E. coli isolates were also classified into phylogenetic groups based on the detection of the chuA, yjaA, and TSPE4.C2 genes. Of the 22 rainwater tanks, 8 (36%) and 5 (23%) were positive for the eaeA (belonging to enteropathogenic E. coli [EPEC] and Shiga-toxigenic E. coli [STEC]) and ST1 (belonging to enterotoxigenic E. coli [ETEC]) genes, respectively. VGs (cdtB, cvaC, ibeA, kpsMT allele III, PAI, papAH, and traT) belonging to extraintestinal pathogenic E. coli (ExPEC) were detected in 15 (68%) of the 22 rainwater tanks. Of the 22 samples, 17 (77%) and 11 (50%) contained E. coli belonging to phylogenetic groups A and B1, respectively. Similarly, 10 (45%) and 16 (72%) contained E. coli belonging to phylogenetic groups B2 and D, respectively. Of the 96 of the 200 strains from 22 tanks that were VG positive, 40 (42%) were carrying a single VG, 36 (37.5%) were carrying two VGs, 17 (18%) were carrying three VGs, and 3 (3%) had four or more VGs. This study reports the presence of multiple VGs in E. coli strains belonging to the STEC, EPEC, ETEC, and ExPEC pathotypes in rainwater tanks. The public health risks associated with potentially clinically significant E. coli in rainwater tanks should be assessed, as the water is used for drinking and other, nonpotable purposes. It is recommended that rainwater be disinfected using effective treatment procedures such as filtration, UV disinfection, or simply boiling prior to drinking.