Current and Future Flow Cytometry Applications Contributing to Antimicrobial Resistance Control

Măruţescu, Luminiţa

doi:10.3390/microorganisms11051300

Cited by 7 publications

(3 citation statements)

References 113 publications

(161 reference statements)

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“…Flow cytometry has become a valuable tool in sepsis research, particularly for quantifying HLA-DR on monocytes, a key marker of immunosuppression in critically ill patients. This technique not only correlates with the ability to mount an effective immune response but also aids in distinguishing sepsis from other conditions like SIRS and in understanding the chronic and treatment-resistant aspects of sepsis [ 30 , 280 ]. The journey towards effective molecular targeted therapies in sepsis has been challenging.…”

Section: Sepsis Therapy: Phenotyping and Personalized Approachesmentioning

confidence: 99%

Advances and Challenges in Sepsis Management: Modern Tools and Future Directions

Santacroce,

D’Angerio,

Ciobanu

et al. 2024

Cells

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Sepsis, a critical condition marked by systemic inflammation, profoundly impacts both innate and adaptive immunity, often resulting in lymphopenia. This immune alteration can spare regulatory T cells (Tregs) but significantly affects other lymphocyte subsets, leading to diminished effector functions, altered cytokine profiles, and metabolic changes. The complexity of sepsis stems not only from its pathophysiology but also from the heterogeneity of patient responses, posing significant challenges in developing universally effective therapies. This review emphasizes the importance of phenotyping in sepsis to enhance patient-specific diagnostic and therapeutic strategies. Phenotyping immune cells, which categorizes patients based on clinical and immunological characteristics, is pivotal for tailoring treatment approaches. Flow cytometry emerges as a crucial tool in this endeavor, offering rapid, low cost and detailed analysis of immune cell populations and their functional states. Indeed, this technology facilitates the understanding of immune dysfunctions in sepsis and contributes to the identification of novel biomarkers. Our review underscores the potential of integrating flow cytometry with omics data, machine learning and clinical observations to refine sepsis management, highlighting the shift towards personalized medicine in critical care. This approach could lead to more precise interventions, improving outcomes in this heterogeneously affected patient population.

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Section: Sepsis Therapy: Phenotyping and Personalized Approachesmentioning

confidence: 99%

Advances and Challenges in Sepsis Management: Modern Tools and Future Directions

Santacroce,

D’Angerio,

Ciobanu

et al. 2024

Cells

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“…For instance, due to the small size of microbes, it is often challenging to differentiate between bacteria and cell debris (Marcos-Fernandez et al 2022). In addition, there are other limitations of non-specific binding of fluorescent dyes, lack of availability of specific antibodies and high cost of antibody-based fluorophores (Iyengar and Robinson 2024; Koch and Muller 2018; Marcos-Fernandez et al 2022; Marutescu 2023; Robinson et al 2023), Cell detection by measuring fluorescence signals (from staining) are limited by the number of lasers and detectors available in some cytometers (Robinson 2019; Robinson et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Measuring autofluorescence spectral signatures for detecting antibiotic-resistant bacteria using Thermofisher’s Bigfoot spectral flow cytometer

Iyengar,

Patsekin,

Rajwa

et al. 2024

Preprint

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Application of flow cytometry to microbiology has been limited due to inadequate availability of bacterial-specific stains, expensive antibody-based fluorophores, ineffective stain cell permeability and challenges in differentiating bacterial cells from cell debris due to their similarity and their small size. Moreover, conventional staining methods require multiple washing steps, limiting sensitivity due to bacterial cells' significantly smaller volume compared to eukaryotic cells. Additionally, most flow cytometers are ill-equipped to handle pathogenic organisms. However, autofluorescence-based detection offers a promising alternative. By leveraging the inherent fluorescence of bacterial cells, the need for extensive washing steps is circumvented, reducing both time and cost. Research indicates that bacterial autofluorescence is primarily associated with specific metabolic components such as Flavin Adenine Dinucleotide (FAD) and Nicotinamide Adenine Dinucleotide (NAD).This study introduces a novel approach to differentiate between clinically isolated antibiotic-resistant and non-resistant bacteria using their autofluorescence spectral signatures. Employing the Bigfoot spectral cytometer, uniquely outfitted with a biosafety cabinet and multiple lasers and fluorescence detectors, bacterial autofluorescence signatures were captured. In a proof-of-concept experiment, E. coli and Salmonella sp. were subjected to gentamicin antibiotics, resulting in increased autofluorescence at distinct excitation and emission wavelengths compared to non-stressed bacteria. Similarly, Methicillin-resistant and susceptible Staphylococcus aureus (MRSA and MSSA) stressed with oxacillin exhibited differential autofluorescence responses, with MSSA showing increased autofluorescence after exposure to oxacillin, while MRSA did not display such changes. These findings suggest that antibiotic-resistant strains can be rapidly identified by monitoring changes in autofluorescence signatures within hours. The label-free, quantitative, and resistant-specific autofluorescence signatures obtained from the Bigfoot spectral flow cytometer hold promise for rapid detection of antibiotic-resistant strains.

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“…An early initiation of empiric antibiotic therapy helps reduce morbidity, mortality, and medical expenses [ 3 ]. Antimicrobial resistance (AMR) poses a growing threat to public health [ 4 ], resulting in 3.57 million deaths worldwide in 2019 [ 5 ]. Consequently, antibiotic choices become more difficult as AMR becomes more prevalent [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

Prevalence and Clinical Characteristics of Bacterial Pneumonia in Neurosurgical Emergency Center Patients: A Retrospective Study Spanning 13 Years at a Tertiary Center

Yang

Yao

et al. 2023

Microorganisms

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Patients with brain injuries are at a heightened susceptibility to bacterial pneumonia, and the timely initiation of empiric antibiotic treatment has been shown to substantially reduce mortality rates. Nevertheless, there is a need for knowledge regarding the resistance and prevalence of pulmonary bacterial infections in this patient population. To address this gap, a retrospective study was conducted at a neurosurgical emergency center, focusing on patients with brain injuries. Among the entire patient population, a total of 739 individuals (18.23%) were identified as having bacterial pneumonia, consisting of 1489 strains of Gram-negative bacteria and 205 strains of Gram-positive bacteria. The resistance of Klebsiella pneumoniae to imipenem exhibited a significant increase, rising from 21.74% in 2009 to 96.67% in 2018, and subsequently reaching 48.47% in 2021. Acinetobacter baumannii displayed resistance rates exceeding 80.0% against multiple antibiotics. The resistance profile of Pseudomonas aeruginosa was relatively low. The proportion of Staphylococcus aureus reached its peak at 18.70% in 2016, but experienced a decline to 7.83% in 2021. The abundance of Gram-negative bacteria exceeded that of Gram-positive bacteria by a factor of 5.96. Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus are prominent pathogens characterized by limited antibiotic choices and scarce treatment alternatives for the isolated strains.

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Current and Future Flow Cytometry Applications Contributing to Antimicrobial Resistance Control

Cited by 7 publications

References 113 publications

Advances and Challenges in Sepsis Management: Modern Tools and Future Directions

Advances and Challenges in Sepsis Management: Modern Tools and Future Directions

Measuring autofluorescence spectral signatures for detecting antibiotic-resistant bacteria using Thermofisher’s Bigfoot spectral flow cytometer

Prevalence and Clinical Characteristics of Bacterial Pneumonia in Neurosurgical Emergency Center Patients: A Retrospective Study Spanning 13 Years at a Tertiary Center

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