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      The microcolony detection method (MCD), a simple and rapid screening test for antimicrobial resistance bacteria on positive blood cultures

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          Abstract

          Rapid detection of antibiotic-resistant bacteria in blood cultures is critical for the timely treatment of patients with sepsis. The aim of this study was to develop a simple method for the rapid detection of drug-resistant bacteria from blood cultures and to evaluate its performance. We developed an optical microscopy-based microcolony detection method (MCD) for the rapid detection of antibiotic-resistant bacterial colonies in media. This method was tested using staphylococci resistant to methicillin and gram-negative bacilli resistant to third-generation cephalosporins and carbapenem. The results of the investigations of clinical samples using this method were compared with the drug susceptibility testing results for each of the 457 isolates, which included 134 staphylococci and 323 g-negative bacilli. The MCD was successful in detecting antibiotic-resistant bacterial growth from culture-positive blood samples in approximately 3 h. The sensitivity/specificity for methicillin-resistant staphylococci was 100%/97.2%. In the case of gram-negative bacilli, the sensitivity/specificity values for bacteria resistant to ceftriaxone, ceftazidime, and carbapenem were 100%/98.7%, 100%/89.3%, and 100%/90.9%, respectively. Therefore, MCD is a clinically useful screening method for the efficient and rapid detection of antibiotic-resistant bacteria and can be easily implemented in laboratories.

          Abstract

          Biotechnology; Microbiology; Genetics; Bacteria; Microbial genomics; Microorganism; Microbial biotechnology; Bloodstream infection; Antimicrobial therapy; Microcolony detection method; MRSA; Extended spectrum β-lactamase production in Enterobacterales; Carbapenem-resistant Enterobacterales.

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          Carbapenemases: the versatile beta-lactamases.

          Anne Marie Queenan, Karen Bush (2007)
          Carbapenemases are beta-lactamases with versatile hydrolytic capacities. They have the ability to hydrolyze penicillins, cephalosporins, monobactams, and carbapenems. Bacteria producing these beta-lactamases may cause serious infections in which the carbapenemase activity renders many beta-lactams ineffective. Carbapenemases are members of the molecular class A, B, and D beta-lactamases. Class A and D enzymes have a serine-based hydrolytic mechanism, while class B enzymes are metallo-beta-lactamases that contain zinc in the active site. The class A carbapenemase group includes members of the SME, IMI, NMC, GES, and KPC families. Of these, the KPC carbapenemases are the most prevalent, found mostly on plasmids in Klebsiella pneumoniae. The class D carbapenemases consist of OXA-type beta-lactamases frequently detected in Acinetobacter baumannii. The metallo-beta-lactamases belong to the IMP, VIM, SPM, GIM, and SIM families and have been detected primarily in Pseudomonas aeruginosa; however, there are increasing numbers of reports worldwide of this group of beta-lactamases in the Enterobacteriaceae. This review updates the characteristics, epidemiology, and detection of the carbapenemases found in pathogenic bacteria.
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            Development of a set of multiplex PCR assays for the detection of genes encoding important beta-lactamases in Enterobacteriaceae.

            Caroline Dallenne, Anaelle da Costa, Dominique Decré … (2010)
            To develop a rapid and reliable tool to detect by multiplex PCR assays the most frequently widespread beta-lactamase genes encoding the OXA-1-like broad-spectrum beta-lactamases, extended-spectrum beta-lactamases (ESBLs), plasmid-mediated AmpC beta-lactamases and class A, B and D carbapenemases. Following the design of a specific group of primers and optimization using control strains, a set of six multiplex PCRs and one simplex PCR was created. An evaluation of the set was performed using a collection of 31 Enterobacteriaceae strains isolated from clinical specimens showing a resistance phenotype towards broad-spectrum cephalosporins and/or cephamycins and/or carbapenems. Direct sequencing from PCR products was subsequently carried out to identify beta-lactamase genes. Under optimized conditions, all positive controls confirmed the specificity of group-specific PCR primers. Except for the detection of carbapenemase genes, multiplex and simplex PCR assays were carried out using the same PCR conditions, allowing assays to be performed in a single run. Out of 31 isolates selected, 22 strains produced an ESBL, mostly CTX-M-15 but also CTX-M-1 and CTX-M-9, SHV-12, SHV-5, SHV-2, TEM-21, TEM-52 and a VEB-type ESBL, 6 strains produced a plasmid-mediated AmpC beta-lactamase (five DHA-1 and one CMY-2) and 3 strains produced both an ESBL (two SHV-12, one CTX-M-15) and a plasmid-mediated AmpC beta-lactamase (DHA-1). We report here the development of a useful method composed of a set of six multiplex PCRs and one simplex PCR for the rapid screening of the most frequently encountered beta-lactamases. This method allowed direct sequencing from the PCR products.
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              Overall burden of bloodstream infection and nosocomial bloodstream infection in North America and Europe.

              M Goto, M.N. Al-Hasan (2013)
              In this systematic review, we estimated the total number of episodes of bloodstream infection (BSI) and deaths from BSI per year in North America and Europe, using data from population-based settings. Then, we estimated the number of episodes and deaths from nosocomial BSI from population-based studies and nosocomial infection surveillance systems. We estimated 575 000-677 000 episodes of BSI per year in North America (536 000-628 000 in the USA and 40 000-49 000 in Canada) and 79 000-94 000 deaths (72 000-85 000 in the USA and 7000-9000 in Canada), using estimates from three population-based studies. We estimated over 1 200 000 episodes of BSI and 157 000 deaths per year in Europe, using estimates from one population-based study in each of the following countries: Denmark (9100 episodes and 1900 deaths), Finland (8700 episodes and 1100 deaths) and England (96 000 episodes and 12 000-19 000 deaths). There were substantial differences in estimates of nosocomial BSI between population-based and nosocomial infection surveillance data. BSI has a major impact on the morbidity and mortality of the general population, as it ranks among the top seven causes of death in all included countries in North America and Europe. However, it is difficult to obtain precise estimates of nosocomial BSI, owing to the limited number of studies. This review highlights the need for a greater focus on BSI research in order to reduce the overall burden of disease by improving the outcome of patients with BSI. It also emphasizes the role of infection control and prevention methods in reducing the burden of nosocomial BSI. ©2013 The Authors Clinical Microbiology and Infection ©2013 European Society of Clinical Microbiology and Infectious Diseases.
              Article 0 comments Cited 232 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Tomokazu Kuchibiro
                Journal
                Journal ID (nlm-ta): Heliyon
                Journal ID (iso-abbrev): Heliyon
                Title: Heliyon
                Publisher: Elsevier
                ISSN (Electronic): 2405-8440
                Publication date PMC-release: 12 November 2020
                Publication date Collection: November 2020
                Publication date (Electronic): 12 November 2020
                Volume: 6
                Issue: 11
                Electronic Location Identifier: e05494
                Affiliations
                [a ]Department of Clinical Laboratory, Naga Municipal Hospital, 1282 Uchita, Kinokawa, Wakayama, 649-6414, Japan
                [b ]Department of Medical Technology and Sciences Facility of Health Sciences, Kyoto Tachibana University, 34 Ohtakuyamadacho, Yamashina-ku, Kyoto City, Kyoto, 607-8175, Japan
                Author notes
                [∗ ]Corresponding author. tk_kensa@ 123456nagahp.jp
                Article
                Publisher Item ID: S2405-8440(20)32337-9 Publisher ID: e05494
                DOI: 10.1016/j.heliyon.2020.e05494
                PMC ID: 7672289
                SO-VID: da63315d-c044-469b-adae-55b4c91b8edc
                Copyright © © 2020 The Author(s)
                License:

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                Date received : 22 June 2020
                Date revision received : 26 June 2020
                Date accepted : 9 November 2020
                Categories
                Subject: Research Article

                Keywords: biotechnology,microbiology,genetics,bacteria,microbial genomics,microorganism,microbial biotechnology,bloodstream infection,antimicrobial therapy,microcolony detection method,mrsa,extended spectrum β-lactamase production in enterobacterales,carbapenem-resistant enterobacterales
                Data availability:
                Keywords: biotechnology, microbiology, genetics, bacteria, microbial genomics, microorganism, microbial biotechnology, bloodstream infection, antimicrobial therapy, microcolony detection method, mrsa, extended spectrum β-lactamase production in enterobacterales, carbapenem-resistant enterobacterales

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