Polymerase chain reaction for the specific detection of Escherichia coli/Shigella.
The outer membrane protein PhoE of members of the family Enterobacteriaceae consists of conserved membrane-spanning segments and hypervariable surface-exposed regions. Two oligonucleotides based on DNA sequences encoding two different cell-surface-exposed regions of the Escherichia coli K12 PhoE protein were tested for their specificity in polymerase chain reactions. They reacted with all strains of the species E. coli/Shigella tested, except for strain S. boydii serovar 13, which is known to represent a different DNA-relatedness group. The probes did not react with any other Enterobacteriaceae tested, including strains of Escherichia blattae, Escherichia hermanii, Escherichia vulneris and Escherichia adecarboxylata, except for an Escherichia fergusonnii strain, which is most closely related to E. coli. Therefore, the primer couple showed a high degree of species-specificity. In addition, a second primer couple based on two conserved regions of the phoE genes was tested. This primer couple recognized a broad group of closely related enteric bacteria including Salmonella and Shigella.
Spierings
G
G
Department of Molecular Cell Biology, Utrecht University, The Netherlands.
Ockhuijsen
C
C
Hofstra
H
H
Tommassen
J
J
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004755:Enterobacteriaceae / Q000235:genetics / Q000302:isolation & purification; D004925:Escherichia / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012760:Shigella / Q000235:genetics / Q000302:isolation & purification*
D001425:Bacterial Outer Membrane Proteins; D015342:DNA Probes; D009841:Oligonucleotides