Abstract: The test was aimed of confirmed these genes distribution condition in positive biofilm with 25 strains of Escherichia coli, and then provided the theoretical basis of the mechanism of biofilm during the pathopoiesia in the Escherichia coli. The polymerase chain reaction was used of being checked virulence genes in the fimH, KpsMTII, chuA, fliC, yja and PapC in 25 strains of Escherichia coli which were positive biofilm. The results showed that, Firstly, the gene of KpsMTII aroused frequence was higjhest which account of 40%; Secondly that the gene of fliC account of 32.5% and the gene of fimH account of 20%, the two genes total was 52.5%; the gene of chuA account of 7.5 %, the genes of yja and PapC unchecked in this test. In this test the biofilm formed were firstly effected in adhere structure genes such as fimH and fliC. Secondly effected gene was KpsMTII, Finally, effected gene was outer membrane protein gene chuA in 25 strains Escherichia coli.