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Selective Uropathogenic E. coli Detection Using Crossed Surface-Relief Gratings.

ABSTRACT: Urinary tract infections (UTIs) are one of the major burdens on public healthcare worldwide. One of the primary causes of UTIs is the invasion of the urinary tract by uropathogenic Escherichia coli (UPEC). Improper treatment of bacterial infections like UTIs with broad-spectrum antibiotics has contributed to the rise of antimicrobial resistance, necessitating the development of an inexpensive, rapid and accurate detection of UPEC. Here, we present real-time, selective and label-free detection of UPEC using crossed surface-relief gratings (CSRGs) as nanometallic sensors incorporated into an optical sensing platform. CSRGs enable real-time sensing due to their unique surface plasmon resonance (SPR)-based light energy exchange, resulting in detection of a very-narrow-bandwidth SPR signal after the elimination of residual incident light. The platform's sensing ability is experimentally demonstrated by the detection of bulk refractive index (RI) changes, with a bulk sensitivity of 382.2 nm/RIU and a resolution in the order of 10^-6 RIU. We also demonstrate, for the first time, CSRG-based real-time selective capture and detection of UPEC in phosphate-buffered saline (PBS) solution, in clinically relevant concentrations, as opposed to other UTI-causing Gram-negative bacteria. The platform's detection limit is calculated to be 10⁵ CFU/mL (concentration on par with the clinical threshold for UTI diagnosis), with a dynamic range spanning four orders of magnitude. This work paves the way for the development of inexpensive point-of-care diagnosis devices focusing on effective treatment of UTIs, which are a burden on public healthcare due to the rise in the number of cases and their recurrences in the recent past.

SUBMITTER: Nair S

PROVIDER: S-EPMC6263983 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Most urinary tract infections (UTIs) are caused by uropathogenic Escherichia coli (UPEC), which depends on an extracellular organelle (type 1 pili) for adherence to bladder cells during infection. Type 1 pilus expression is partially regulated by inversion of a piece of DNA referred to as fimS, which contains the promoter for the fim operon encoding type 1 pili. fimS inversion is regulated by up to five recombinases collectively known as Fim recombinases. These Fim recombinases are currently known to regulate two other switches: the ipuS and hyxS switches. A long-standing question has been whether the Fim recombinases regulate the inversion of other switches, perhaps to coordinate expression for adhesion or virulence. We answered this question using whole-genome sequencing with a newly developed algorithm (structural variation detection using relative entropy [SVRE]) for calling structural variations using paired-end short-read sequencing. SVRE identified all of the previously known switches, refining the specificity of which recombinases act at which switches. Strikingly, we found no new inversions that were mediated by the Fim recombinases. We conclude that the Fim recombinases are each highly specific for a small number of switches. We hypothesize that the unlinked Fim recombinases have been recruited to regulate fimS, and fimS only, as a secondary locus; this further implies that regulation of type 1 pilus expression (and its role in gastrointestinal and/or genitourinary colonization) is important enough, on its own, to influence the evolution and maintenance of multiple additional genes within the accessory genome of E. coli IMPORTANCE UTI is a common ailment that affects more than half of all women during their lifetime. The leading cause of UTIs is UPEC, which relies on type 1 pili to colonize and persist within the bladder during infection. The regulation of type 1 pili is remarkable for an epigenetic mechanism in which a section of DNA containing a promoter is inverted. The inversion mechanism relies on what are thought to be dedicated recombinase genes; however, the full repertoire for these recombinases is not known. We show here that there are no additional targets beyond those already identified for the recombinases in the entire genome of two UPEC strains, arguing that type 1 pilus expression itself is the driving evolutionary force for the presence of these recombinase genes. This further suggests that targeting the type 1 pilus is a rational alternative nonantibiotic strategy for the treatment of UTI.

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Selective Uropathogenic E. coli Detection Using Crossed Surface-Relief Gratings.

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