Identification of Pseudomonas aeruginosa on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms.
In the present study, 40 clinical strains of Pseudomonas aeruginosa were investigated by using the polymerase chain reaction (PCR). We used primers to amplify spacer regions between the 16S and 23S genes in the prokaryotic rRNA genetic loci. When the spacer amplification products were resolved by electrophoresis, the resulting patterns were characteristic for all tested strains. Only one specific PCR fragment of 580 bp was formed. This product was digested with HaeIII, HinfI and AluI restriction endonucleases. Restriction fragments produced by all three restriction endonucleases were characteristic and have the same sizes for all strains tested. The amplification product contains a conserved, internal single HaeIII restriction site. On the basis of our results, PCR amplifications of the 16S-23S spacer region for Pseudomonas aeruginosa and subsequent RFPL analysis show significant promise as a tool for the simple identification of this bacteria.
Department of Microbiology, Technical University of Gdansk, Poland.
J
J
Kur
A
A
Burkiewicz
A
A
Zablocka
E
E
Gospodarek
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S