The Sysmex UF-5000 was used as recommended by the company (Sysmex Corporation). It uses a flowcytometry-based system, with forward scatter light (FSC), side scatter light (SSC) and side fluorescent light (SFL). In addition to these components, the Sysmex UF-5000 uses depolarized side scattered light (DSS) to differentiate between red blood cells and crystals. The machine is fully automated, has a modular concept for urinalysis workflow and can be used with UD-10, a newly developed urine image viewer. The samples were analyzed on automatic mode with the recommended long rinse cycle switched on to avoid carryover between samples [11 ]. The bact. info flag provides information about the type of bacteria detected in the sample (Gram negative, Gram positive, combinations of the two or unspecified) based on different light signals of FSC, SFL and SHH for Gram positive and Gram negative bacteria, due to different dye intake by the cell wall structures.
Uf 5000
Manufactured by Sysmex
Sourced in Japan
The UF-5000 is a fully automated urine cell analyzer that provides comprehensive analysis of urine samples. It employs flow cytometry technology to perform a detailed examination of cellular components, including red blood cells, white blood cells, and other particles, in the urine. The UF-5000 generates accurate and reliable data to support clinical decision-making processes.
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Lab products found in correlation
Uc 3500, by Sysmex (2 mentions)
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Uriselect 4 agar, by Bio-Rad (1 mentions)
Vitek 2 system, by bioMérieux (1 mentions)
Au5800, by Beckman Coulter (1 mentions)
Xn 3000, by Sysmex (1 mentions)
Cobas 8000, by Roche (1 mentions)
Behring nephelometer 2 analyzer, by Siemens (1 mentions)
Uf 1000i, by Sysmex (1 mentions)
18 protocols using uf 5000
1
Automated Bacterial Identification in Urine
2
Automated Urine Analysis with Sysmex UF-5000
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Following inoculation, we analysed the urine samples for all parameters available in the Sysmex UF-5000 instrument (Sysmex), which is the latest urine and body fluid analyser from Sysmex. In the instrument, cells are counted and classified by analysing forward scattered light, side scattered light, side fluorescent light and depolarized side scattered light (Sysmex).
Sysmex UF-5000 has a theoretical maximum throughput of 105 urine samples per hour, assuming no rinsing between samples (Sysmex). Automated rinsing between samples can be programmed to reduce the risk of sample-to-sample carryover from samples with a high bacterial count to samples with a lower bacterial count [11 (link)]. In this study we programmed rinsing according to the manufacturer’s instructions with rinse mode 0-0-1-3-5 as follows: no rinsing after samples with ≤9.9×105 BACT ml−1, one rinse after samples with 1×106–9.9×106 BACT ml−1, three rinses after samples with 1×107–9.9×107 BACT ml−1, and five rinses after samples with ≥108 BACT ml−1 (Table 2 ). Sample throughput will therefore depend of the number of rinses, which again depends on the concentration of bacteria in the samples.
Sysmex UF-5000 has a theoretical maximum throughput of 105 urine samples per hour, assuming no rinsing between samples (Sysmex). Automated rinsing between samples can be programmed to reduce the risk of sample-to-sample carryover from samples with a high bacterial count to samples with a lower bacterial count [11 (link)]. In this study we programmed rinsing according to the manufacturer’s instructions with rinse mode 0-0-1-3-5 as follows: no rinsing after samples with ≤9.9×105 BACT ml−1, one rinse after samples with 1×106–9.9×106 BACT ml−1, three rinses after samples with 1×107–9.9×107 BACT ml−1, and five rinses after samples with ≥108 BACT ml−1 (
3
Rapid Cell Count Comparison
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Effluent samples of unknown cell concentrations, from randomly selected patients were collected from 13 patients without peritonitis, 9 patients with suspected peritonitis, and 7 patients with peritonitis on antibiotic therapy within 4 hours of sample drain. Samples were received on separate days and measured independently. Patient effluent was added to cuvettes and measured simultaneously on the QuickCheck device, and using a Sysmex UF-5000, as previously. To prevent bias, cell counts on each cell counting device were performed blind by independent operators. Each sample was measured in triplicate and average data shown. Linearity of the data was determined for QuickCheck and Sysmex UF-5000.
4
Automated Urine Flow Cytometry Analysis
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The Sysmex UF-5000 (Sysmex Corporation, Kobe, Japan) is a fully automated urine flow cytometry analyzer with the newest technology, using a blue semiconductor laser at a 488 nm wavelength, with a surface channel (SFch) and a core channel (CRch) analysis chamber [11 ]. Like its predecessor, the Sysmex UF-5000 analyzer has technology for particle counting and classification based on signals of forward-scattered light (FSC), side-scattered light (SSC), side fluorescent light (SFL), and depolarized side-scattered light (DSS). It determines bacteria using a different light signal for FSC, SFL, and SSC for Gram-negatives and Gram-positives based on different dye intake by cell wall structures. The flagging that appears in the Sysmex UF-5000 analyzer can be BACT: Gram Negative?, BACT: Gram Positive?, and BACT: Gram Pos/Neg? A minimum volume of 2.0 mL of uncentrifuged urine is needed in automated mode and 0.6 mL in the STAT mode. In both the modes, the aspiration volume is 0.45 mL [11 ].
5
Urine Specific Gravity Measurement
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Specific gravity (S.G., mass of a unit volume) is the density ratio of urine with respect to water. It was measured with Sysmex UF-5000 (SYSMEX CORPORATION, Kobe, Japan).
6
Automated Urinalysis Using Sysmex UF-5000
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UFC analysis was performed by the Sysmex UF-5000 (Sysmex Corporation, Kobe, Japan) with an automated rinse (settings 0,1,1,7,7) after samples with a high bacterial count to avoid carryover [22 (link)]. Analysis results of bacterial count per µL (BACT/µL), WBC count per µL (WBC/µL), squamous epithelial cell count per µL, and analysis time were recorded from the Sysmex UF-5000. Analysis was performed by either the project assistants or lab workers trained to use the Sysmex UF-5000. Only results with a time from urine collection to analysis between zero and two hours were used for our calculations. The urine culture results and the expert panel diagnosis were not available at the time of analysis.
7
Isolation and Enumeration of Leukocytes
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Leukocytes were isolated from PD effluent samples by centrifugation at 400g for 5 minutes. Supernatants were discarded and any red blood cells removed using 5 ml dH2O (30 s), followed by neutralization with 5 ml of 1.8% NaCl (Sigma-Aldrich). Jurkat cells and clinically-derived leukocytes were both subjected to centrifugation at 400g for 5 minutes and cell pellets resuspended in 5 ml of 0.22 μm filtered effluent, counted using flow cytometry (Sysmex UF-5000) (Sysmex UK Ltd, Milton Keynes, UK), and diluted in filtered effluent to 1 × 104 cells/μl. Test cell concentrations were prepared by serial dilution in effluent to produce 10, 25, 50, 75, 100, 500, and 1 × 103 cells/μl.18 For each cell counting method, measurements were taken simultaneously and performed blind by independent operators in a randomized order. Further details are provided in the Supplementary Methods .
8
Automated Urinalysis with Sysmex UF-5000
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The Sysmex UF-5000 was used according to the manufacturer’s instructions, as given by the company (Sysmex Corporation). The Sysmex UF-5000 uses a flowcytometry-based system, with forward scatter light (FSC), side scatter light (SSC), and side fluorescent light (SFL). In addition to these components, the Sysmex UF-5000 uses depolarized side scattered light (DSS) to differentiate between red blood cells and crystals. The machine is fully automated, has a modular concept for urinalysis workflow, and can be used.
9
Evaluation of Urine Tests for UTI Diagnosis
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The midstream urine specimens sent to the clinical microbiology laboratory of our hospital for urine culture between September 13, 2021, and November 15, 2021, were collected. The urine specimens were collected in a sterile urine cup for urine culture first, and the remaining urine specimens were simultaneously tested by the URIT 1600 chemical analyzer (Youlite Corp, Guilin, China) and Sysmex UF-5000 (Sysmex, Kobe, Japan). The time interval between urine culture and analyses was less than 6 hours. Urine culture was used to ascertain UTI in these patients, and those with urine bacteria ≥104 CFU /ml were defined as UTI patients, according to the standards of previous studies [12 (link), 13 (link)]. Urine nitrate, WBC, bacteria, urine culture results, and the Gram flag were used for analysis. Patients aged <18 years were excluded from this study. The authors had access to information that could identify individual participants during data collection.
10
Urine Sediment Analysis Protocols
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All test data were processed qualitatively and analyzed statistically, and manual microscopic examination was used as the gold standard. Positive criteria were RBC >3 p/HPF, WBC >5 p/HPF, and cast >1 p/LPF.7 , 8 Therefore, the positive criteria after Sysmex UF‐5000 system conversion were RBC >0.54 p/μl, WBC >0.9 p/μl, and cast >2.9 p/μl. The positive criteria after Atellica UAS 800 analyzer conversion were RBC >13.2 p/μl, WBC >22 p/μl, and cast >10.7 p/μl. The qualitative results of the sediment components in the selected urine samples were compared by the coincidence rate and the false‐negative rate. Coincidence rate = 100% × [(a + d) /n], false‐negative rate = 100% × [c/(a + c)]. Kappa value was calculated by Kappa test in SPSS 25.0 statistical software for the evaluation of the consistency of the results of manual microscopy. Kappa scale values ranges were 0.41–0.60, moderate consistency; 0.61–0.80, high consistency; and 0.81–1, complete consistency.
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