Application of biotinylated and 32P probes for detection of P-fimbriae in urinary E. coli
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P-fimbriae, a common cause of urinary tract infections in children, were detected using DNA probes. The P gene was prevalent in pyelonephritis cases, less so in symptomatic UTIs, and absent in asymptomatic bacteriuria.
Area of Science:
- Microbiology
- Molecular Biology
- Pediatrics
Background:
- Escherichia coli (E. coli) is a primary pathogen causing urinary tract infections (UTIs).
- P-fimbriae are virulence factors associated with E. coli pathogenicity in UTIs.
- Urinary tract infections in children range from asymptomatic bacteriuria to severe pyelonephritis.
Purpose of the Study:
- To investigate the prevalence of P-fimbriae in E. coli strains isolated from children with different UTI classifications.
- To evaluate the efficacy of biotinylated and 32P-DNA probes for detecting the P gene in E. coli.
Main Methods:
- Isolation of 26 E. coli strains from pediatric patients with pyelonephritis, symptomatic UTIs, and asymptomatic bacteriuria.
- Preparation of biotinylated and 32P-DNA probes specific for P-fimbriae genes.
- Hybridization assays using the prepared DNA probes to detect the presence of the P gene in E. coli isolates.
Main Results:
- Eleven of 13 E. coli strains from pyelonephritis patients were positive for the P gene.
- Only 3 of 9 E. coli strains from symptomatic UTI cases harbored the P gene.
- No E. coli strains from asymptomatic bacteriuria cases tested positive for the P gene.
- The biotinylated probe demonstrated high applicability in standard laboratory settings.
Conclusions:
- The P gene, indicative of P-fimbriae, is strongly associated with pyelonephritis in pediatric E. coli infections.
- P-fimbriae are less common in symptomatic UTIs and absent in asymptomatic bacteriuria.
- Biotinylated DNA probes offer a practical and reliable method for detecting P-fimbriae in diagnostic laboratories.
Abstract:
Escherichia coli is the common causative agent of urinary tract infections. Twenty-six strains of Escherichia coli were isolated from children with pyelonephritis, symptomatic urinary tract infections and asymptomatic bacteriuria. Biotinylated and 32P-DNA probes were prepared for detection of P-fimbriae in the isolates. Of the 13 strains isolated from patients with pyelonephritis 11 were positive for the presence of the P gene by both probes. Strains isolated from cases of symptomatic urinary tract infections revealed the presence of P gene only in three samples of the total of nine isolated. None of the isolated E. coli strains from asymptomatic bacteriuria was found positive for the presence of the P gene. The biotinylated probe was simple and easily applicable in standard laboratory conditions and therefore the authors recommend it for use in diagnostic laboratories.