ObjectiveTo explore the genetic marks of continuously isolated Escherichia coli (E.coli) and Pseudomonas aeruginosa (P. aeruginosa) and the correlation with nosocomial infection (NI). MethodsPolymerase chain reaction (PCR)  was used to detect plasmid AmpC genes (ACT1 and DHA), class Ⅰ integrase gene (IntⅠ1) and disinfectantresistant gene (qacE△1sul1) of E.coli; Four kinds of genetic marks of  integron and transposon (qacE△1sul1, merA, tnpA, tnpU) in P. aeruginosa were also  detected by PCR.  ResultsAmong E.coli, the positive rate of plasmid AmpC genes of ACT1 and DHA was 57.50% and 40.00% respectively, the positive rate of IntⅠ1 and qacE△1 sul1 was 47.50% and 57.50% respectively. Among P. aeruginosa, the positive rate of qacE△1sul1 and merA  was 48.57%  and 11.43% respectively. There were neither gene tnpA nor gene tnpU  in P. aeruginosa strains. ConclusionE.coli isolated from the clinic carries plasmid  AmpC genes, IntⅠ1 genes and qacE△1sul1; P.aeruginosa carries qacE△1sul1 and merA. Resistant genes among different bacteria strains can be transmitted, which is significant in epidemiology.