Restriction fragment length polymorphism of PCR amplified papE gene products is correlated with complete serotype among uropathogenic Escherichia coli isolates.
Using whole bacteria, rather than extracted, purified DNA samples, we amplified the papE gene sequences from 63 Escherichia coli isolates belonging to O serogroups O1, O2 and O6. These isolates were from collections separated temporally as well as geographically: from four cities in the US and one in Sweden. PCR amplified papE products were digested with restriction endonucleases and the relative sizes of the fragments compared for each strain. For 41 of the strains, we found a correlation between the papE restriction fragment length polymorphism (RFLP) and the complete serotype. Furthermore, we were able to detect the presence of duplicate copies of the gene in 14 of the isolates; these 14 isolates were among the 22 that did not exhibit a correlation between the RFLP of their amplified papE sequences and their complete serotype. We conclude that RFLP analysis of PCR products is a rapid and relatively simple method for examining the DNA of E. coli containing the pap gene sequence.
Jantausch
B A
BA
Children's National Medical Center, George Washington University Medical Center, Washington, DC 20010, USA.
Hull
S I
SI
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002648:Child; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D004927:Escherichia coli Infections / Q000382:microbiology; D029968:Escherichia coli Proteins; D005260:Female; D010861:Fimbriae, Bacterial / Q000737:chemistry / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D006801:Humans; D007223:Infant; D008565:Membrane Proteins; D005810:Multigene Family / Q000235:genetics; D009693:Nucleic Acid Hybridization; D019081:O Antigens / Q000276:immunology; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D006180:Proton-Translocating ATPases; D012756:Sheep; D013548:Sweden; D014481:United States; D014552:Urinary Tract Infections / Q000382:microbiology
D001426:Bacterial Proteins; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D019081:O Antigens; D006180:Proton-Translocating ATPases; C101709:AtpH protein, E coli