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. 2021 Jul 6;16(7):e0254064.
doi: 10.1371/journal.pone.0254064. eCollection 2021.

Rapid diagnosis and reduced workload for urinary tract infection using flowcytometry combined with direct antibiotic susceptibility testing

Hanne Margrethe Gilboe  1 , Olaug Marie Reiakvam  1 , Linda Aasen  1 , Trygve Tjade  1 , Johan Bjerner  1 , Trond Egil Ranheim  1 , Peter Gaustad  1
Affiliations

Rapid diagnosis and reduced workload for urinary tract infection using flowcytometry combined with direct antibiotic susceptibility testing

Hanne Margrethe Gilboe et al. PLoS One. .

Abstract

Background: We evaluated if flowcytometry, using Sysmex UF-5000, could improve diagnosis of urinary tract infections by rapid identification of culture negative and contaminated samples prior to culture plating, thus reducing culture plating workload and response time. We also evaluated if it is possible to reduce the response time for antibiotic susceptibility profiles using the bacteria information flag on Sysmex UF-5000 to differentiate between Gram positive and negative bacteria, followed by direct Antibiotic Susceptibility Testing (dAST) on the positive urine samples.

Methods: One thousand urine samples were analyzed for bacteria, white blood cells and squamous cells by flowcytometry before culture plating. Results from flowcytometric analysis at different cut-off values were compared to results of culture plating. We evaluated dAST on 100 urine samples that were analyzed as positive by flowcytometry, containing either Gram positive or Gram negative bacteria.

Results: Using a cut-off value with bacterial count ≥100.000/mL and WBCs ≥10/μL, flowcytometry predicted 42,1% of samples with non-significant growth. We found that most contaminated samples contain few squamous cells. For 52/56 positive samples containing Gram negative bacteria dAST was identical to routine testing. Overall, there was concordance in 555/560 tested antibiotic combinations.

Conclusion: Flowcytometry offers advantages for diagnosis of urinary tract infections. Screening for negative urine samples on the day of arrival reduces culture plating and workload, and results in shorter response time for the negative samples. The bacteria information flag predicts positive samples containing Gram negative bacteria for dAST with high accuracy, thus Antibiotic Susceptibility Profile can be reported the day after arrival. For the positive samples containing Gram negative bacteria the concordance was very good between dAST and Antibiotic Susceptibility Testing in routine. For positive samples containing Gram positive bacteria the results were not convincing. We did not find any correlation between epithelial cells and contamination.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Flowcytometric findings (bact./mL) in culture plated with significant growth (significant findings) and non-significant growth (non-significant findings).
Fig 2
Fig 2. Flowcytometric findings (WBCs/μl) in culture plated samples with significant growth (significant findings) and non-significant growth (non-significant findings).
Fig 3
Fig 3. Number of squamous cells in contaminated samples.
Fig 4
Fig 4. Algorithm of UTI diagnosis using flowcytometry with reported samples on different days.
See this image and copyright information in PMC

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