Flow cytometric analysis of viable bacteria in urine samples of febrile patients at the emergency department

Tavenier, Anne H.; Boer, Foppie J de; Moshaver, Bijan; Leur, Sjef J C M van der; Stegeman, Coen A.; Groeneveld, P. H. P.

doi:10.1002/cyto.b.21548

Cited by 9 publications

(8 citation statements)

References 28 publications

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“…The combination of fluorescent cell permeable and impermeable nucleic acid probes and light scattering properties of bacterial particles allows for the viability assessment of bacterial communities Calcein violet-acetoxymethyl ester (CV-AM) has been tested to measure the susceptibility of Mycobacterium tuberculosis to a variety of bacteriostatic antibiotics and was found to be reliable [46]. Flow cytometry with a propidium iodide-SYTO costaining technique has been used for rapid detection of urinary tract infections, and was found to have a sensitivity of 100% and specificity of 78.4% [47,48]. This dual staining technique with propidium iodide, a viability assessment fluorophore, offers the potential for assessing environmental bacteria for resistance to bactericidal antibiotics.…”

Section: Flow Cytometry As a High-throughput Culture Alternativementioning

confidence: 99%

Antibiotic Resistance in Minimally Human-Impacted Environments

Scott

Lee

2020

IJERPH

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Antibiotic resistant bacteria (ARB) have become contaminants of concern in environmental systems. Studies investigating environmental ARB have primarily focused on environments that are greatly impacted by anthropogenic activity. Background concentrations of ARB in natural environments is not well understood. This review summarizes the current literature on the monitoring of ARB and antibiotic resistance genes (ARGs) in environments less impacted by human activity. Both ARB and ARGs have been detected on the Antarctic continent, on isolated glaciers, and in remote alpine environments. The methods for detecting and quantifying ARB and ARGs from the environment are not standardized and warrant optimization. Further research should be focused on the detection and quantification of ARB and ARGs along human gradients to better characterize the factors leading to their dissemination in remote environments.

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Section: Flow Cytometry As a High-throughput Culture Alternativementioning

confidence: 99%

Antibiotic Resistance in Minimally Human-Impacted Environments

Scott

Lee

2020

IJERPH

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“…Another reason for disagreement could be due to the possible counting of nonviable bacteria by the Sysmex UF-5000. Although technical solutions to separate non-viable from viable bacteria using special dyes in UFC exist in certain instruments [20,21], this is not included as an option in the Sysmex UF-5000. Thus, urine samples from pregnant women should always be cultured, for the detection of uropathogens as well as group B streptococci [22].…”

Section: Discussionmentioning

confidence: 99%

Use of Sysmex UF-5000 flow cytometry in rapid diagnosis of urinary tract infection and the importance of validating carryover rates against bacterial count cut-off

Haugum

Haugan

Skage

et al. 2021

Journal of Medical Microbiology

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Introduction. Urinary tract infections are common bacterial infections worldwide. Urine culture is the gold standard method to identify and quantify the presence or absence of bacteria in urine. Flow cytometry, which can differentiate and quantify multiple particles (including bacteria) in the urine, presents an alternative method for rapid screening to rule out bacteriuria. Hypothesis. Adding flow cytometry to identify urine samples without bacteriuria could substantially reduce the number of urine samples that need to be cultured as well as the response time for negative results. However, the level of instrument rinsing between samples could affect sample-to-sample carryover rate, a concept given little attention in previous studies. Aim. We aimed to evaluate urine flow cytometry as a rapid screening method to identify urine samples without significant bacterial growth, including analyses of cross-contamination and sample-to-sample carryover rate. Methodology. We analysed 3919 urine samples by quantitative urine culture and flow cytometry screening (Sysmex UF-5000). Receiver operator characteristic (ROC) curve analyses were used to test method agreement to identify: (a) positive vs. negative culture and (b) mixed vs. pure culture. In addition, we performed carryover and cross-contamination studies. Results. ROC curve analyses identified bacterial count (BACT ml−1) and leucocyte count (WBC µl−1) as possible predictors of bacterial growth in the total material and subpopulations, except pregnant women (n=451). This subgroup was excluded from further analyses, leaving a final 3468 urine samples. Area under the ROC curve was 0.94 (95 % CI 0.93–0.95) and 0.81 (95 % CI 0.79–0.82) for bacterial and leucocyte count, respectively. A bacterial count cut-off of 30 BACT ml−1 resulted in 95.2 % sensitivity and 91.2 % negative predictive value, resulting in approximately 30 % of urine samples that could be reported as negative without culture. Use of high-level rinse modes was necessary to ensure carryover rates <0.05 %. Conclusion. Flow cytometry is a suitable and rapid method to rule out urine samples without significant bacterial growth. Rinses between samples should be adjusted, depending on the cut-off used, to prevent sample-to-sample carryover, whereas cross-contamination can be eliminated by the use of separate urine aliquots for flow cytometry analysis and urine culturing respectively.

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“…A set of specialized equipment and experimental methods that are currently used to develop new antibiotics and antiviral compounds includes, among others, such techniques as flow cytometry [1], spectroscopy [2], fluorometry [3], scanning electron microscopy (SEM) [4], and transmission electron microscopy (TEM) [5]. Recently, atomic force microscopy (AFM) found its place as a technology with Pathogens 2020, 9, 969 2 of 30 great potential to study the microorganisms themselves as well as the molecules that might control their growth.…”

Section: Application Of Atomic Force Microscopy (Afm) In the Field Of Microbiologymentioning

confidence: 99%

Physics Comes to the Aid of Medicine—Clinically-Relevant Microorganisms through the Eyes of Atomic Force Microscope

et al. 2020

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Despite the hope that was raised with the implementation of antibiotics to the treatment of infections in medical practice, the initial enthusiasm has substantially faded due to increasing drug resistance in pathogenic microorganisms. Therefore, there is a need for novel analytical and diagnostic methods in order to extend our knowledge regarding the mode of action of the conventional and novel antimicrobial agents from a perspective of single microbial cells as well as their communities growing in infected sites, i.e., biofilms. In recent years, atomic force microscopy (AFM) has been mostly used to study different aspects of the pathophysiology of noninfectious conditions with attempts to characterize morphological and rheological properties of tissues, individual mammalian cells as well as their organelles and extracellular matrix, and cells’ mechanical changes upon exposure to different stimuli. At the same time, an ever-growing number of studies have demonstrated AFM as a valuable approach in studying microorganisms in regard to changes in their morphology and nanomechanical properties, e.g., stiffness in response to antimicrobial treatment or interaction with a substrate as well as the mechanisms behind their virulence. This review summarizes recent developments and the authors’ point of view on AFM-based evaluation of microorganisms’ response to applied antimicrobial treatment within a group of selected bacteria, fungi, and viruses. The AFM potential in development of modern diagnostic and therapeutic methods for combating of infections caused by drug-resistant bacterial strains is also discussed.

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Flow cytometric analysis of viable bacteria in urine samples of febrile patients at the emergency department

Cited by 9 publications

References 28 publications

Antibiotic Resistance in Minimally Human-Impacted Environments

Antibiotic Resistance in Minimally Human-Impacted Environments

Use of Sysmex UF-5000 flow cytometry in rapid diagnosis of urinary tract infection and the importance of validating carryover rates against bacterial count cut-off

Physics Comes to the Aid of Medicine—Clinically-Relevant Microorganisms through the Eyes of Atomic Force Microscope

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