A total of 161 Escherichia coli (E. coli) strains isolated from children with urinary tract infection (UTI) were analysed for the genes encoding the virulence factors such as pyelonephritis (pap), s fimbriae (sfa), afimbrial adhesin I (afaI), haemolysin (hly), cytotoxic necrotising factor I (cnf I) and aerobactin (aer) by multiplex PCR. Ninety-four E. coli strains were found to carry at least one virulence factor. Therefore, 58.38% of total population was positive for one virulence gene at least. Percentage of genes within the total population for pap, sfa, afaI, hly, cnf I and aer was found as 22.98, 6.21, 9.94, 1.24, 9.94 and 39.75, respectively. Our analysis showed that sfa-pap (p < 0.001); pap-aer, afaI-aer and cnf I-pap (P < 0.05) and hly-sfa (p < 0.01) significantly co-occurred in their respective samples. In the light of these findings, we suggest an important role of pap causing UTI.