Culture-free bacterial detection and identification from blood with rapid, phenotypic, antibiotic susceptibility testing

Shi, Xuyang; Kadiyala, Usha; Yau, S.‐T.

doi:10.1038/s41598-018-21520-9

Cited by 31 publications

(26 citation statements)

References 27 publications

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“…The Field Effect Enzymatic Detection (FEED) biosensor platform can detect extremely low bacterial concentrations (below 10 c.f.u./ml). An electrical field between the working electrode and the immune complex multiplies the biocatalytic output current, enabling a direct detection of bacteria without sample processing (Shi et al, 2018 ). These biosensors apply horse radish peroxidase (HRP) as a redox source in the sandwich hybridization complex.…”

Section: Proof-of-concept Technologiesmentioning

confidence: 99%

Modern Tools for Rapid Diagnostics of Antimicrobial Resistance

Vasala

Hytönen

Laitinen

2020

Front. Cell. Infect. Microbiol.

209

146

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Fast, robust, and affordable antimicrobial susceptibility testing (AST) is required, as roughly 50% of antibiotic treatments are started with wrong antibiotics and without a proper diagnosis of the pathogen. Validated growth-based AST according to EUCAST or CLSI (European Committee on Antimicrobial Susceptibility Testing, Clinical Laboratory Standards Institute) recommendations is currently suggested to guide the antimicrobial therapy. Any new AST should be validated against these standard methods. Many rapid diagnostic techniques can already provide pathogen identification. Some of them can additionally detect the presence of resistance genes or resistance proteins, but usually isolated pure cultures are needed for AST. We discuss the value of the technologies applying nucleic acid amplification, whole genome sequencing, and hybridization as well as immunodiagnostic and mass spectrometry-based methods and biosensor-based AST. Additionally, we evaluate the potential of integrated systems applying microfluidics to integrate cultivation, lysis, purification, and signal reading steps. We discuss technologies and commercial products with potential for Point-of-Care Testing (POCT) and their capability to analyze polymicrobial samples without pre-purification steps. The purpose of this critical review is to present the needs and drivers for AST development, to show the benefits and limitations of AST methods, to introduce promising new POCT-compatible technologies, and to discuss AST technologies that are likely to thrive in the future.

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Section: Proof-of-concept Technologiesmentioning

confidence: 99%

Modern Tools for Rapid Diagnostics of Antimicrobial Resistance

Vasala

Hytönen

Laitinen

2020

Front. Cell. Infect. Microbiol.

209

146

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“…Methods using molecular biology and genetics can deliver AST results within 3 h but only enable the detection of known antibiotic resistance genes (Ellington et al, 2017 ; She and Bender, 2018 ). Early detection by culture-based methods coupled with optical microscopy, video imaging, micro-fluidic approaches, immunoassays, bio-sensors, and machine learning (Mohan et al, 2013 ; Kelley, 2017 ; Shi et al, 2018 ), yielding results within 3 to 6 h (Le Page et al, 2015 ; Choi et al, 2017 ; Smith et al, 2017 ), have been reported and are proposed as a possible way to replace routinely used methods. Other common AST methods used in clinical laboratories are broth micro-dilution, disk diffusion and E-test methods (Baker et al, 1991 ).…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Imipenem Resistance in Gram-Negative Bacteria Using Tabletop Scanning Electron Microscopy: A Preliminary Evaluation

et al. 2021

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Background: Enabling faster Antimicrobial Susceptibility Testing (AST) is critical, especially to detect antibiotic resistance, to provide rapid and appropriate therapy and to improve clinical outcomes. Although several standard and automated culture-based methods are available and widely used, these techniques take between 18 and 24 h to provide robust results. Faster techniques are needed to reduce the delay between test and results.Methods: Here we present a high throughput AST method using a new generation of tabletop scanning electron microscope, to evaluate bacterial ultra-structural modifications associated with susceptibilities to imipenem as a proof of concept. A total of 71 reference and clinical strains of Gram-negative bacteria were used to evaluate susceptibility toward imipenem after 30, 60, and 90 min of incubation. The length, width and electron density of bacteria were measured and compared between imipenem susceptible and resistant strains.Results: We correlated the presence of these morphological changes to the bacterial susceptibility and their absence to the bacterial resistance (e.g., Pseudomonas aeruginosa length without [2.24 ± 0.61 μm] and with [2.50 ± 0.68 μm] imipenem after 30 min [p = 3.032E-15]; Escherichia coli width without [0.92 ± 0.07 μm] and with [1.28 ± 0.19 μm] imipenem after 60 min [p = 1.242E-103]). We validated our method by a blind test on a series of 58 clinical isolates where all strains were correctly classified as susceptible or resistant toward imipenem.Conclusion: This method could be a potential tool for rapidly identifying carbapenem-resistance in Enterobacterales in clinical microbiology laboratories in <2 h, allowing the empirical treatment of patients to be rapidly adjusted.

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“…Approximately 30-50% of patients receive ineffective antibiotic therapy because physicians must treat immediately with first-line, broad-spectrum antibiotics until the results of culture-based detection are available [3]. For example, blood culture assays require 48 to 72 h to complete [4], with fastidious organisms such as Bacillus species and HACEK (Haemophilus species, Aggregatibacter species, Cardiobacterium hominis, Eikenella corrodens, and Kingella species) organisms requiring several days to produce a conclusive result [5]. This not only affects patient survival due to the prescription of inappropriate antibiotics [6,7] but the misuse of antibiotics is a direct contributor to the global spread of antimicrobial resistant bacteria [8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Reporter Phage-Based Detection of Bacterial Pathogens: Design Guidelines and Recent Developments

Meile

Kilcher

Loessner

et al. 2020

Viruses

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Fast and reliable detection of bacterial pathogens in clinical samples, contaminated food products, and water supplies can drastically improve clinical outcomes and reduce the socio-economic impact of disease. As natural predators of bacteria, bacteriophages (phages) have evolved to bind their hosts with unparalleled specificity and to rapidly deliver and replicate their viral genome. Not surprisingly, phages and phage-encoded proteins have been used to develop a vast repertoire of diagnostic assays, many of which outperform conventional culture-based and molecular detection methods. While intact phages or phage-encoded affinity proteins can be used to capture bacteria, most phage-inspired detection systems harness viral genome delivery and amplification: to this end, suitable phages are genetically reprogrammed to deliver heterologous reporter genes, whose activity is typically detected through enzymatic substrate conversion to indicate the presence of a viable host cell. Infection with such engineered reporter phages typically leads to a rapid burst of reporter protein production that enables highly sensitive detection. In this review, we highlight recent advances in infection-based detection methods, present guidelines for reporter phage construction, outline technical aspects of reporter phage engineering, and discuss some of the advantages and pitfalls of phage-based pathogen detection. Recent improvements in reporter phage construction and engineering further substantiate the potential of these highly evolved nanomachines as rapid and inexpensive detection systems to replace or complement traditional diagnostic approaches.

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Culture-free bacterial detection and identification from blood with rapid, phenotypic, antibiotic susceptibility testing

Cited by 31 publications

References 27 publications

Modern Tools for Rapid Diagnostics of Antimicrobial Resistance

Modern Tools for Rapid Diagnostics of Antimicrobial Resistance

Rapid Detection of Imipenem Resistance in Gram-Negative Bacteria Using Tabletop Scanning Electron Microscopy: A Preliminary Evaluation

Reporter Phage-Based Detection of Bacterial Pathogens: Design Guidelines and Recent Developments

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