Assuntos
Infecções Urinárias/microbiologia , Urina/microbiologia , Escherichia coli Uropatogênica/isolamento & purificação , beta-Lactamases/genética , beta-Lactamases/metabolismo , Idoso , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Análise por Conglomerados , Feminino , Genes Bacterianos , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Tipagem Molecular , Reação em Cadeia da Polimerase , República da Coreia , Análise de Sequência de DNA , Escherichia coli Uropatogênica/classificação , Escherichia coli Uropatogênica/enzimologia , Escherichia coli Uropatogênica/genética , Resistência beta-LactâmicaRESUMO
A total of 100 clinical isolates of Escherichia coli (n = 35), Klebsiella pneumoniae (n = 63), Proteus mirabilis (n = 1), and Salmonella serovar Stanley (n = 1), showing resistance to cefoxitin, or returning positive in extended-spectrum ß-lactamase (ESBL) by Clinical and Laboratory Standards Institute (CLSI) ESBL confirmatory method, were studied. The isolates were examined by the boronic acid (BA) disk test, polymerase chain reaction, and pulsed-field gel electrophoresis (PFGE) to investigate genetic similarities. The concurrence rates for ESBLs by the CLSI and the BA disk test were 97% for E. coli and 96.7% for K. pneumoniae. A total of 41 isolates showing cefoxitin resistance yielded all positive by the BA disk test. All the 33 K. pneumoniae isolates, which showed positive by the BA disk test, were carrying AmpC genes. The TEM and CTX-M types were predominant in E. coli and the SHV and the CIT and/or DHA types were predominant in K. pneumoniae. PFGE analysis showed almost 75% of genetic similarities among K. pneumoniae isolates producing ESBLs and/or AmpC ß-lactamases (AmpCs) as each K. pneumoniae carried variable genes and showed variable antibiotic patterns. Clearly, the BA disk test was a useful method for the detection of ESBLs and AmpCs. In particular, cefoxitin resistance and BA-positive trait of K. pneumoniae do reflect the presence of AmpC genes in the organism.
Assuntos
Proteínas de Bactérias/metabolismo , Escherichia coli/enzimologia , Klebsiella pneumoniae/enzimologia , Proteus mirabilis/enzimologia , Salmonella/enzimologia , beta-Lactamases/metabolismo , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Farmacorresistência Bacteriana Múltipla , Eletroforese em Gel de Campo Pulsado , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/isolamento & purificação , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Prevalência , Proteus mirabilis/efeitos dos fármacos , Proteus mirabilis/genética , Proteus mirabilis/isolamento & purificação , República da Coreia , Salmonella/efeitos dos fármacos , Salmonella/genética , Salmonella/isolamento & purificação , beta-Lactamases/biossíntese , beta-Lactamases/genéticaRESUMO
Various tyrosine phosphorylation motif regions of H. pylori cagA exist. The number of these regions was found to have some influence on cell signaling, which was found to be more pronounced when in D (ESS) region than in C (WSS) region. A molecular biological method with multiplex PCR was developed to distinguish C and D regions, and to identify the repetition number of tyrosine phosphorylation of the cagA gene. Multiplex PCR using novel primer sets was performed on 73 strains of H. pylori isolated from Korean patients with upper gastrointestinal diseases. The Western cagA was identified in only 3 strains (4.1%) whereas East Asia cagA was identified in 69 strains (94.5%). These results were reconfirmed through a sequencing analysis. The method developed in this study would be useful for monitoring the repeated number of C and D regions of tyrosine phosphorylation motifs in H. pylori cagA.