Ultrafast and Multiplexed Bacteriophage Susceptibility Testing by Surface Plasmon Resonance and Phase Imaging of Immobilized Phage Microarrays

O’Connell, Larry; Mandula, Ondřej; Leroy, Loïc; Aubert, Axelle; Marcoux, Pierre R.; Roupioz, Yoann

doi:10.3390/chemosensors10050192

Cited by 10 publications

(10 citation statements)

References 44 publications

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“…The targeted, addressable immobilization of phages on the sensor surface is required to apply surface plasmon resonance for phage susceptibility testing, and this task is far from simple 132,133 . Under prior findings of earlier studies, the purification and immobilization process utilized in this work consistently produces homogenous, high-purity and highdensity phage monolayers from suspensions of the "phage gh-1" and "phage44AHJD" 131 . In another study, using the fulllength Det7 phage tail protein (Det7T), a surface plasmon resonance biosensor can quickly and accurately identify "Salmonella enterica serovar Typhimurium" (S. Typhimurium) 134 .…”

Section: Surface Plasmon Resonance Biosensorsmentioning

confidence: 66%

“…It has been demonstrated that methods based on "phase imaging" or methods based on "surface plasmon resonance imaging (SPRi)" are possibilities for rapid (less than two hours) phage susceptibility testing in the broth phase. Covalently immobilized arrays of the "phage 44AHJD, "phage P68", and "phage gh-1" were used to create biosensing layers, which were then subjected to liquid cultures of either Pseudomonas putida or methicillin-resistant Staphylococcus aureus (MRSA) 131 . The targeted, addressable immobilization of phages on the sensor surface is required to apply surface plasmon resonance for phage susceptibility testing, and this task is far from simple 132,133 .…”

Section: Surface Plasmon Resonance Biosensorsmentioning

confidence: 99%

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Advancement of Phage Therapy Approaches in The Battle of Multi-Drug Resistance: A Review

Kumari¹,

Chakraborty²,

Ghosh³

2023

Int J Life Sci Pharm Res

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One of the most significant issues faced by humanity today is antibiotic resistance. Drugs are used in such vast amounts for human health, aquatic life, and agricultural animals that harmful bacteria have developed antibiotic resistance to various antibiotics. Further, the usage of antibiotics is increasing because of situations such as increased infections and chronic diseases that need antimicrobial treatment. Since antimicrobial resistance is rising, it is necessary to take action to help reduce and eliminate infectious diseases and ensure animal and human health. Because of this, many attempts are being made to tackle multi-drugresistant bacteria. Among the many advanced techniques that are occurring, the use of phage therapy is one such emerging procedure. The main aim of this prospective review is to identify the various new phage formulations available as a potential therapeutic intervention to combat multidrug resistance among bacteria and the objective is to identify the various reasons associated with the induction of the phenomenon of "multidrug resistance" among different bacteria, focusing on the use of phage therapy, its advantages as well as disadvantages over antibiotics as a possible therapeutic intervention. Various phage formulations, such as phage cocktails with antibiotics, nanoparticles, phage-delivering hydrogels, and many more, are emerging formulations that have successful results in fighting against multi-drug-resistant bacteria. Commercial phage solutions have helped combat antimicrobial resistance in poultry and livestock farms, improving everyone's health worldwide. As a result, this study shall serve as a source of information and understanding of the concerns mentioned above for the entirety of society and every human community.

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Section: Surface Plasmon Resonance Biosensorsmentioning

confidence: 66%

Section: Surface Plasmon Resonance Biosensorsmentioning

confidence: 99%

Advancement of Phage Therapy Approaches in The Battle of Multi-Drug Resistance: A Review

Kumari¹,

Chakraborty²,

Ghosh³

2023

Int J Life Sci Pharm Res

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“…Although these can be collected in high-throughput and in real time, they do not directly measure damage and can therefore be difficult to interpret 13 . Other fast methods, such as the OmniLog™ system and surface plasmon resonance have been adapted to monitor phage infections, but these require specific equipment, making it more challenging to implement their use 31 , 32 . In sum, there is still room for improvement in methods for determining phage sensitivity, as there is currently no clear standard.…”

Section: Discussionmentioning

confidence: 99%

Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye

Egido

Toner-Bartelds

Costa

et al. 2023

Sci Rep

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Bacteriophages (phages) are viruses that specifically attack bacteria. Their use as therapeutics, which constitutes a promising alternative to antibiotics, heavily relies on selecting effective lytic phages against the pathogen of interest. Current selection techniques are laborious and do not allow for direct visualization of phage infection dynamics. Here, we present a method that circumvents these limitations. It can be scaled for high-throughput and permits monitoring of the phage infection in real time via a fluorescence signal readout. This is achieved through the use of a membrane-impermeant nucleic acid dye that stains the DNA of damaged or lysed bacteria and new phage progeny. We have tested the method on Pseudomonas aeruginosa and Klebsiella pneumoniae and show that an increase in fluorescence reflects phage-mediated killing. This is confirmed by other techniques including spot tests, colony plating, flow cytometry and metabolic activity measurements. Furthermore, we illustrate how our method may be used to compare the activity of different phages and to screen the susceptibility of clinical isolates to phage. Altogether, we present a fast, reliable way of selecting phages against Gram-negative bacteria, which may be valuable in optimizing the process of selecting phages for therapeutic use.

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“…For example, Low et al described a method that identifies phage-host interactions by fluorescence tagging of phages with subsequent tracking of the infection using flow cytometry and confocal fluorescence microscopy, along with live/dead assay for phage susceptibility (Low et al, 2020). There are also studies where the prediction of phage infectivity was approached through machine learning and imaging (Perlemoine et al, 2021;Lood et al, 2022;O'Connell et al, 2022). The study by Perlemoine et al introduced a custom wide-field lens-less imaging device, which can be used for phage plaque detection (Perlemoine et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

“…The methodology can detect phage-resistant bacterial micro-colonies within 3 hours, however, the scalability to screen multiple phages has not yet been demonstrated. More recently, a study utilizing surface plasmon resonance imaging (SPRi) and phase imaging for phage susceptibility testing demonstrated phage detection in less than two hours ( O'Connell et al., 2022 ). However, the proof-of-concept study was performed only with Staphylococcus aureus and Pseudomonas putida bacterial species.…”

Section: Introductionmentioning

confidence: 99%

Rapid hydrogel-based phage susceptibility test for pathogenic bacteria

Patpatia

Schaedig

Dirks

et al. 2022

Front. Cell. Infect. Microbiol.

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Phage therapy is one alternative to cure infections caused by antibiotic resistant bacteria. Due to the narrow host range of phages, hundreds to thousands of phages are required to cover the diversity of bacterial pathogens. In personalized phage therapy, fast selection of the phages for individual patients is essential for successful therapy. The aims of this study were to set up a rapid hydrogel-based liquid phage susceptibility assay (PST) for the selection of phages for therapeutic use and to establish a “ready-to-screen” plate concept, where phages are readily stored in hydrogel as small droplets in microtiter plate wells. We first tested four commercially available hydrogels (GrowDex, Askina, Purilon, and Intrasite) for their suitability as phage matrices in PSTs with four phages, two of which infecting Escherichia coli and two Staphylococcus aureus. Of these four hydrogels, GrowDex was the best matrix for PST, as it did not inhibit bacterial growth, released phages quickly when mixed with bacterial culture, and maintained phage viability well. We then optimized the assay for both optical density and microscopy readers using GrowDex as matrix with 23 bacterial strains representing 10 different species and 23 phages possessing different morphologies and genome sizes. When the bacterial growth was monitored by microscopy reader, the PST was executed in just 3 hours, and there was no need for overnight culturing bacterial cells prior to the assay, whereas using optical density reader, bacteria had to be pre-cultured overnight, and the assay time was five hours. Finally, we evaluated the effect of three different chemical stabilizers (trehalose, hyaluronic acid, and gelatin) in a six-month stability assay with six model phages. These phages assay behaved very differently in respect to the chemical stabilizers, and there was not a single stabilizer suitable for all phages. However, when gelatin (0.01%) or hyaluronic acid (0.2 mg/ml) was used as stabilizer, all tested phages were still considered as positives in PST after a six-month storage in 1 ml volume. In “ready-to-screen” plates, the differences in phage stabilities were even more profound, varying from two to six months for the most and least stable phages, respectively.

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Ultrafast and Multiplexed Bacteriophage Susceptibility Testing by Surface Plasmon Resonance and Phase Imaging of Immobilized Phage Microarrays

Cited by 10 publications

References 44 publications

Advancement of Phage Therapy Approaches in The Battle of Multi-Drug Resistance: A Review

Advancement of Phage Therapy Approaches in The Battle of Multi-Drug Resistance: A Review

Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye

Rapid hydrogel-based phage susceptibility test for pathogenic bacteria

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