Abstract
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.
Highlights
AND FOREWORDThere is an unmet need for rapid and decentralized diagnostics in outpatient clinics to reduce the misuse of antibiotics
Standard growth-based technologies based on disc diffusion and broth dilution still dominate in Antimicrobial susceptibility testing (AST)
Few rapid growth-based AST methods work directly with polymicrobial clinical samples, which is required in Point of Care Testing (POCT)
Summary
There is an unmet need for rapid and decentralized diagnostics in outpatient clinics to reduce the misuse of antibiotics. Since immunodetection does not necessarily require disruption of the target microbes, it can potentially provide pathogen identification and growth monitoring in a single step It is applicable as simple lateral flow (LF) tests, but can be integrated to biosensor technology, microfluidics and even to DNA/RNA-based analysis. A light-emitting diode array and fiber-based optics enabled detection of turbidity changes in wells already after 1 min (Feng et al, 2016) This system, tested for 17 antibiotics targeting Gram-negative bacteria on clinical isolates of K. pneumoniae, provided drug susceptibility interpretation with accuracy of 99.23%. The system showed a total categorical agreement of 97.9% with standard disk diffusion assays, but a careful standardization of cell densities prior to cultivation was found necessary (Veses-Garcia et al, 2018)
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