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Evaluation of flow cytometry for cell count and detection of bacteria in biological fluids

N. Dossou, I. Gaubert, C. Moriceau, E. Cornet, S. Le Hello, D. Malandain
doi: https://doi.org/10.1101/2021.10.04.463144
N. Dossou
1CHU de Caen Normandie, Department of Microbiology, 14000 Caen, France
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  • For correspondence: ndossou@hotmail.fr
I. Gaubert
1CHU de Caen Normandie, Department of Microbiology, 14000 Caen, France
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C. Moriceau
2CHU de Caen Normandie, Department of Haematology, 14000 Caen, France
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E. Cornet
2CHU de Caen Normandie, Department of Haematology, 14000 Caen, France
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S. Le Hello
1CHU de Caen Normandie, Department of Microbiology, 14000 Caen, France
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D. Malandain
1CHU de Caen Normandie, Department of Microbiology, 14000 Caen, France
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Abstract

The analysis of biological fluids is crucial for the diagnosis and monitoring of diseases causing effusions and helps in the diagnosis of infectious diseases. The gold standard method for cell count in biological fluids is the manual method using counting chambers. The microbiological routine procedures consist of Direct Gram staining and culture on solid or liquid media. We evaluate the analytical performance of SYSMEX UF4000 (Sysmex, Kobe, Japan) and Sysmex XN10 (Sysmex, Kobe, Japan) in comparison with cytological and microbiological routine procedures. A total of 526 biological fluid samples were included in this study (42 ascitic, 31 pleural, 31 peritoneal, 125 cerebrospinal, 281 synovial, and 16 peritoneal dialysis fluids). All samples were analysed by flow cytometry and subsequently processed following cytological and/or microbiological routine procedures. With regards to cell counts, UF4000 (Sysmex, Kobe, Japan) showed a performance which was at least equivalent to those of the reference methods and superior to those of XN10 (Sysmex, Kobe, Japan). Moreover, the bacterial count obtained with UF4000 (Sysmex, Kobe, Japan) was significantly higher among culture or Direct Gram stain positive samples. We established 3 optimal cut-off points to predict Direct Gram stain positive samples for peritoneal (465.0 bacteria/μL), synovial (1200.0 bacteria/μL), and cerebrospinal fluids (17.2 bacteria/μL) with maximum sensitivity and negative predictive values. Cell count and detection of bacteria by flow cytometry could be used upstream cytological and microbiological routine procedures to improve and accelerate the diagnosis of infection of biological fluid samples.

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Posted October 07, 2021.
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Evaluation of flow cytometry for cell count and detection of bacteria in biological fluids
N. Dossou, I. Gaubert, C. Moriceau, E. Cornet, S. Le Hello, D. Malandain
bioRxiv 2021.10.04.463144; doi: https://doi.org/10.1101/2021.10.04.463144
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Evaluation of flow cytometry for cell count and detection of bacteria in biological fluids
N. Dossou, I. Gaubert, C. Moriceau, E. Cornet, S. Le Hello, D. Malandain
bioRxiv 2021.10.04.463144; doi: https://doi.org/10.1101/2021.10.04.463144

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