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  • In vivo detection of Escherichia coli type 1 fimbrial expression and phase variation during experimental urinary tract infection.

In vivo detection of Escherichia coli type 1 fimbrial expression and phase variation during experimental urinary tract infection.

Microbiology (Reading, England) (1999-10-28)
C Struve, K A Krogfelt
ABSTRACT

Adhesion mediated by fimbriae is thought to play an important role in the pathogenesis of urinary tract infections (UTI) by Escherichia coli. The majority of clinical isolates of E. coli from UTI are able to express type 1 fimbriae. However, the importance of these fimbriae as a virulence factor has been controversial. To investigate the expression of type 1 fimbriae in vivo during UTI, mice were transurethrally infected with uropathogenic E. coli C175-94 and type 1 fimbrial expression was determined directly by two independent methods at 2 h, 1 d and 3 d after infection. By use of an assay combining in situ rRNA hybridization and immunofluorescence, all bacterial cells detected in urine, bladders and kidneys from mice sacrificed 1 and 3 d after onset of infection were found to express type 1 fimbriae. In contrast, the majority of cells in the suspension used for infection of mice and specimens from mice sacrificed 2 h after inoculation were found to be non-fimbriated. Similar results were obtained with a PCR assay revealing the orientation of the invertible promoter driving the transcription of type 1 fimbrial genes. Whilst the promoter in both ON and OFF positions could be amplified from the suspension used for infection and specimens from mice sacrificed 2 h after inoculation, at 1 and 3 d after onset of infection only the promoter in the ON orientation could be amplified. These results show that introduction of E. coli C175-94 into the mouse urinary tract leads to markedly enhanced expression of type 1 fimbriae.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Escherichia coli FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH)
Sigma-Aldrich
Escherichia coli FISH probe -ATTO488, Probe for fluorescence in situ hybridization (FISH), 20µM in water