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      Modern Tools for Rapid Diagnostics of Antimicrobial Resistance

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          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.

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          Most cited references129

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          Analysis of discrimination mechanisms in the mammalian olfactory system using a model nose.

          Olfaction exhibits both high sensitivity for odours and high discrimination between them. We suggest that to make fine discriminations between complex odorant mixtures containing varying ratios of odorants without the necessity for highly specialized peripheral receptors, the olfactory systems makes use of feature detection using broadly tuned receptor cells organized in a convergent neurone pathway. As a test of this hypothesis we have constructed an electronic nose using semiconductor transducers and incorporating design features suggested by our proposal. We report here that this device can reproducibly discriminate between a wide variety of odours, and its properties show that discrimination in an olfactory system could be achieved without the use of highly specific receptors.
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            Nucleic acid sequence-based amplification.

            J. Compton (1991)
            Nucleic acid sequence-based amplification (NASBA) is a primer-dependent technology that can be used for the continuous amplification of nucleic acids in a single mixture at one temperature.
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              EUCAST expert rules in antimicrobial susceptibility testing.

              EUCAST expert rules have been developed to assist clinical microbiologists and describe actions to be taken in response to specific antimicrobial susceptibility test results. They include recommendations on reporting, such as inferring susceptibility to other agents from results with one, suppression of results that may be inappropriate, and editing of results from susceptible to intermediate or resistant or from intermediate to resistant on the basis of an inferred resistance mechanism. They are based on current clinical and/or microbiological evidence. EUCAST expert rules also include intrinsic resistance phenotypes and exceptional resistance phenotypes, which have not yet been reported or are very rare. The applicability of EUCAST expert rules depends on the MIC breakpoints used to define the rules. Setting appropriate clinical breakpoints, based on treating patients and not on the detection of resistance mechanisms, may lead to modification of some expert rules in the future. © 2011 The Authors. Clinical Microbiology and Infection © 2011 European Society of Clinical Microbiology and Infectious Diseases.
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                Author and article information

                Contributors
                Journal
                Front Cell Infect Microbiol
                Front Cell Infect Microbiol
                Front. Cell. Infect. Microbiol.
                Frontiers in Cellular and Infection Microbiology
                Frontiers Media S.A.
                2235-2988
                15 July 2020
                2020
                : 10
                : 308
                Affiliations
                [1] 1Protein Dynamics, Faculty of Medicine and Health Technology, Tampere University , Tampere, Finland
                [2] 2Fimlab Laboratories , Tampere, Finland
                Author notes

                Edited by: Francois Vandenesch, Université de Lyon, France

                Reviewed by: Jerome Lemoine, Université de Lyon, France; Karsten Becker, University Medicine Greifswald, Germany

                *Correspondence: Antti Vasala antti.i.vasala@ 123456gmail.com

                This article was submitted to Clinical Microbiology, a section of the journal Frontiers in Cellular and Infection Microbiology

                Article
                10.3389/fcimb.2020.00308
                7373752
                32760676
                6352817c-132a-40c6-915a-e345634f9428
                Copyright © 2020 Vasala, Hytönen and Laitinen.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 17 January 2020
                : 22 May 2020
                Page count
                Figures: 2, Tables: 2, Equations: 0, References: 161, Pages: 23, Words: 17976
                Categories
                Cellular and Infection Microbiology
                Review

                Infectious disease & Microbiology
                antibiotic resistance,antimicrobial susceptibility test,antimicrobial resistance,point of care test,rapid ast

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