J Bacteriol Virol.  2017 Mar;47(1):54-63. 10.4167/jbv.2017.47.1.54.

Clonal and Virulence Distribution of Uropathogenic Escherichia coli Isolated from Children in Korea

• Kim DH
• Choi CH

• KMID: 2384420
• DOI: http://doi.org/10.4167/jbv.2017.47.1.54

Abstract

Urinary tract infections (UTI) are one of the most frequent infectious diseases. Uropathogenic Escherichia coli (UPEC) are among major pathogens causing UTI. A variety of virulence genes are mainly responsible for the severity of these emerging infection. This study investigate the influences of virulence properties of UPEC isolates with reference to multi-locus sequence typing (MLST). The aim of this study was targeted that investigation of the bacterial pathogenicity associated with UTI in children. A total of 58 UPEC isolates were collected from urine samples from patients with clinical diagnosis of uncomplicated UTI. The MLST of UPEC strains were assessed by methods based on polymerase chain reaction. Motility was evaluated using soft-agar plates. Biofilm formation was analyzed in microtiter dish biofilm formation assay. Cell death assay was analyzed by Annexin V/Phosphatidylserine staining and DNA fragmentation assay. According the result, the predominant sequence type (ST) was ST95 (24.1%) and ST73 (17.2%). There were some difference in virulence gene and antibiotics resistance between ST95 and ST73. The number of 11 (18.9%) isolates were strongly adherent. Based on the detected biofilm formation, these strongly adherent are almost ST73. The ST95 was higher than ST73 in population, but ST95 was lower than ST73 in motility and cell death induction. This study indicated that the UPEC molecular strains are related to some virulence traits. Furthermore, the virulence factors carried by ST73 strains contribute to their abilities to colonize the host and cause disease.

Keyword

Uropathogenic Escherichia coli; Virulence; MLST; Sequence type; Clonal complex

MeSH Terms

Anti-Bacterial Agents
Biofilms
Cell Death
Child*
Colon
Communicable Diseases
Diagnosis
DNA Fragmentation
Humans
Korea*
Polymerase Chain Reaction
Urinary Tract Infections
Uropathogenic Escherichia coli*
Virulence Factors
Virulence*
Anti-Bacterial Agents
Virulence Factors

Figure

• 

  Figure 1. Biofilm formation of UPEC isolates. UPEC isolates incubate for 24 h at 37°C in LB broth without NaCl. Biofilm was stained by 0.1% crystal violet. Error bars represent the standard deviations of the results from three independent experiments.

• 

  Figure 2. Motility of UPEC. (A and B) Motility on 0.3% LB Agar of CFT073, isolated UPEC No.2, 22, 26-1, 53 and 29 after 16 h of incubation at 37°C. The errors bars in panel A represent the standard deviations from four independent experiments. Significant difference in motility was to determined using one-way ANOVA; ∗, p < 0.005.

• 

  Figure 3. UPEC-induced cell death. (A) Apoptosis of T24 infected with UPEC No. 2, 22, 38, 26-1 or 53 (MOI 10) for 3 h was measured by flow cytometry using Annexin V/PI staining. (B) DNA fragmentation analysis of T24 cells infected with UPEC isolate No. 26-1. T24 cells were infected with UPEC isolate No. 26-1 at an MOI of 10 for different time point at 37°C. M, 100 bp DNA marker; Ctrl, no infection.

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