Rapid, sensitive detection of bacteria in platelet samples with Fountain Flow Cytometry

Johnson, Paul E.; Moriwaki, Mika; Johnson, Joseph Taber

doi:10.1002/jcla.22115

Cited by 2 publications

(2 citation statements)

References 10 publications

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“…Conventional FCM is a cytological instrument which can detect high-volume of cell populations and subpopulations [11,13]. It has been reported that conventional FCM can successfully examine bacteria in blood component and urine [14,15], viruses in seawater [16] and virus-infected cells in blood or bronchoalveolar lavage fluid [17]. It can even detect and quantify airborne fungi [18].…”

Section: Introductionmentioning

confidence: 99%

Application of single-cell real-time imaging flow cytometry in rapid detection of pathogenic fungi in clinical liquid specimens

Dong

Zheng

et al. 2021

Methods Appl. Fluoresc.

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Rapid and direct observation of fungal spores or hyphae in clinical liquid specimens poses a challenge for the diagnosis of invasive fungal infection. To allow rapid detection of fungal pathogens, we designed a new method of fungal cell detection involving double fluorescence staining with calcium fluorescent white (CFW) and SYTOX green combined with single-cell real-time imaging flow cytometry (IFC). IFC allowed quick detection and analysis of detailed morphology of the spores and pseudohyphae of Candida albicans, and small hyphae and typical truncated large mycelia of Aspergillus fumigatus. Further, cell sorting based on fluorescence, the width-to-height ratio and bright-field parameters preferentially identified spores or hyphae with a typical cell wall. The specificity and overall coincidence rate of IFC for fungi detection in common clinical samples were 100% and 98.18%, respectively. Moreover, the detection rate by IFC (102/105, 97.14%) was significantly higher (P = 0.002) than that by wet mount method (89/105, 84.5%). Therefore, IFC is a reliable diagnostic method with a high potential for application for rapid diagnosis of fungal infection in the clinic.

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Section: Introductionmentioning

confidence: 99%

Application of single-cell real-time imaging flow cytometry in rapid detection of pathogenic fungi in clinical liquid specimens

Dong

Zheng

et al. 2021

Methods Appl. Fluoresc.

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“…The real-time PCR is often prone to false-positive results because of DNA contamination of the PCR reagents. , FCM is also used for the detection of PC contaminations. While effective, several modifications including culturing of PCs in bacterial growth media are needed for low concentration levels of bacteria in platelets, which limits the detection capability. − Other obstacles for FCM include slow-growing bacteria not reaching titers at high enough levels to be detected because of differential growth kinetics. Some of these methods are also used for the testing of blood cultures for bloodstream infection detection. ,,, Hence, new technologies are arguably needed that can detect pathogens in simple and complex media with minimal sample preparation, without the need for further sample amplification, and with low turnaround time.…”

Section: Introductionmentioning

confidence: 99%

Multimodal Magneto-Fluorescent Nanosensor for Rapid and Specific Detection of Blood-Borne Pathogens

Banerjee

Tummala

Elliott

et al. 2019

ACS Appl. Nano Mater.

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Detection of bacterial contaminants in blood and platelet concentrates (PCs) continues to be challenging in clinical settings despite available current testing methods. At the same time, it is important to detect the low bacterial contaminants present at the time of transfusion. Herein, we report the design and synthesis of a dual-modal magnetofluorescent nanosensor (MFnS) by integrating magnetic relaxation and fluorescence modalities for the wide-range detection of blood-borne pathogens. In this study, functional MFnSs are designed to specifically detect Staphylococcus epidermidis and Escherichia coli, two of the predominant bacterial contaminants of PCs. Specific interaction between the target pathogen and functional MFnS resulted in the change of water proton's magnetic relaxation time, indicative of sensitive detection of the target bacteria from low to high colony-forming units (CFUs). In addition, the acquired magnetic relaxation signal of the MFnS further facilitated quantitative assessment of the slow and fast growth kinetics of target pathogens. Moreover, the presence of fluorescence modality in MFnS allowed for the detection of multicontaminants. Bacterial detection was also performed in complex media including whole blood and PCs, which further demonstrated its robust detection sensitivity. Overall, our study indicated that the designer MFnS will have potential for the wide-range detection of blood-borne pathogens and features desirable qualities including timeliness, sensitivity, and specificity.

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Rapid, sensitive detection of bacteria in platelet samples with Fountain Flow Cytometry

Abstract: This pilot study supports the efficacy of FFC for the rapid and sensitive screening of platelet donations for bacteria.

Cited by 2 publications

References 10 publications

Application of single-cell real-time imaging flow cytometry in rapid detection of pathogenic fungi in clinical liquid specimens

Application of single-cell real-time imaging flow cytometry in rapid detection of pathogenic fungi in clinical liquid specimens

Multimodal Magneto-Fluorescent Nanosensor for Rapid and Specific Detection of Blood-Borne Pathogens

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