Rectal Carriage of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae in Hospitalized Patients: Selective Preenrichment Increases Yield of Screening

Bergh, Marjolein Kluytmans-van den; Verhulst, Carlo; Willemsen, Linette E. M.; Verkade, Erwin; Bonten, Marc J. M.; Kluytmans, Jan

doi:10.1128/jcm.01251-15

Cited by 55 publications

(38 citation statements)

References 21 publications

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“…(), we used a semi‐selective enrichment step prior to plating on selective ESBL agar, which may have affected the recovery rate. The difference in the performance of semi‐selective (using broth selecting for Gram negative rods) and selective enrichment (i.e., broth additionally containing a third‐generation cephalosporin) remains to be elucidated and comparative data are limited (Blane et al., ; Jazmati, Hein, & Hamprecht, ; Kluytmans‐Van Den Bergh et al., ).…”

Section: Antimicrobial Resistant Escherichia Coli Obtained By Nonselementioning

confidence: 99%

Antimicrobial resistant and extended‐spectrum β‐lactamase producing Escherichia coli in common wild bird species in Switzerland

et al. 2019

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A total of 294 fecal swabs from 294 wild birds in Switzerland were cultivated for antimicrobial resistant (AMR) Escherichia coli. Samples were also subcultivated to detect E. coli with extended‐spectrum β‐lactamases (ESBL), carbapenemases, and plasmid‐mediated aminoglycoside or colistin resistance, respectively. Samples from 17 (5.8%) of the birds contained 19 AMR E. coli, whereof 26.3% were multidrug resistant. Five (1.7%) ESBL‐producing E. coli were detected. The isolates harbored bla CTX‐M‐1 (two isolated from carrion crows and from one great spotted woodpecker, respectively), bla CTX‐M‐15 (originating from a grey heron), bla CTX‐M‐55 (from a carrion crow), and bla CTX‐M‐65 (from a common blackbird). Phylogenetic analysis assigned three isolates to commensal phylogroups A and B1, one to extraintestinal pathogenic group B2, and one to phylogroup F. Multilocus sequence typing identified sequence types (STs) that have been found previously in ESBL E. coli in wild birds (ST58, ST205, ST540). One isolate harboring bla CTX‐M‐55 was assigned to the recently emerged fluoroquinolone‐resistant, extraintestinal pathogenic E. coli clone ST1193. Wild birds have the potential to disperse AMR, including clinically important resistance genes, from anthropogenic‐influenced habitats to diverse areas, including vulnerable natural environments such as surface waters or mountain regions.

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Section: Antimicrobial Resistant Escherichia Coli Obtained By Nonselementioning

confidence: 99%

Antimicrobial resistant and extended‐spectrum β‐lactamase producing Escherichia coli in common wild bird species in Switzerland

et al. 2019

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“…Detection of ESBL-E in clinical cultures was performed according to national and international guidelines (29,30). Screening cultures consisted of perianal swabs that were preenriched in a selective tryptic soy broth and subsequently cultured on a selective ESBL screening agar (EbSA) (Cepheid Benelux, Apeldoorn, the Netherlands) as described previously (31). For all oxidase-negative Gram-negative bacteria that grew on the EbSA plate, species identification was performed with Vitek MS (bioMérieux, Marcy l'Etoile, France), using the Vitek MS Knowledge Base v1.1 (until January 2013) or v2.0 (from January 2013).…”

Section: Esbl-e Isolatesmentioning

confidence: 99%

Whole-Genome Multilocus Sequence Typing of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae

et al. 2016

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e Molecular typing has become indispensable in the detection of nosocomial transmission of bacterial pathogens and the identification of sources and routes of transmission in outbreak settings, but current methods are labor-intensive, are difficult to standardize, or have limited resolution. Whole-genome multilocus sequence typing (wgMLST) has emerged as a whole-genome sequencing (WGS)-based gene-by-gene typing method that may overcome these limitations and has been applied successfully for several species in outbreak settings. In this study, genus-, genetic-complex-, and species-specific wgMLST schemes were developed for Citrobacter spp., the Enterobacter cloacae complex, Escherichia coli, Klebsiella oxytoca, and Klebsiella pneumoniae and used to type a national collection of 1,798 extended-spectrum-beta-lactamase-producing Enterobacteriaceae (ESBL-E) isolates obtained from patients in Dutch hospitals. Genus-, genetic-complex-, and species-specific thresholds for genetic distance that accurately distinguish between epidemiologically related and unrelated isolates were defined for Citrobacter spp., the E. cloacae complex, E. coli, and K. pneumoniae. wgMLST was shown to have higher discriminatory power and typeability than in silico MLST. In conclusion, the wgMLST schemes developed in this study facilitate high-resolution WGS-based typing of the most prevalent ESBL-producing species in clinical practice and may contribute to further elucidation of the complex epidemiology of antimicrobial-resistant Enterobacteriaceae. wgMLST opens up possibilities for the creation of a Web-accessible database for the global surveillance of ESBL-producing bacterial clones. The continuing global spread of extended-spectrum-beta-lactamase-producing Enterobacteriaceae (ESBL-E) constitutes a major public health threat in both health care and community settings (1-4). Extended-spectrum beta-lactamases (ESBL) confer resistance to the majority of beta-lactam antibiotics, including third-generation cephalosporins, which limits the options for antimicrobial therapy and results in increased morbidity and mortality and health care costs (5, 6). Infection control guidelines recommend prevention of the spread of ESBL-E in health care settings (7,8). Molecular typing has become an important tool in infection control, as it enables the detection of nosocomial transmission of bacterial pathogens and the identification of sources and routes of transmission in outbreak settings. Different molecular typing methods have been used, ranging from fingerprintbased methods, like pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP), to sequencebased methods, like multilocus sequence typing (MLST) (9-11). Although widely adopted, PFGE is labor-intensive and difficult to standardize and has limited interlaboratory reproducibility (12)(13)(14). AFLP is less time-consuming than PFGE but may have lower discriminatory power for typing of Enterobacteriaceae (13, 15). MLST targets 7 or 8 housekeeping genes, depending on the Ent...

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“…A more recent report from the Netherlands showed that Ͼ25% of the ESBL-E carriers may only be identified by preenrichment when a rectal swab was used (12). Rectal swabs are believed to be inadequate for the recovery and detection of various pathogens due to high variation in the quantity of fecal material on each swab and the low inoculum (13).…”

mentioning

confidence: 99%

Use of an Enrichment Broth Improves Detection of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae in Clinical Stool Samples

2016

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cThis study evaluated the impact of preenrichment on the detection of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E) in clinical stool samples. ESBL-E were detected in 41 of 343 patients (12.0%). As 31.7% of the ESBL-E carriers were identified by preenrichment, only this additional diagnostic step significantly improved the detection of ESBL-E. E xtended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E) are increasingly reported worldwide. Treatment options for infections due to these organisms are limited, and initial empirical therapy is often inappropriate and is associated with increased morbidity and mortality (1). ESBL-E carriers are known to develop bacteremia with the isolates colonizing the patients' intestines (2). Therefore, in many hospitals, risk-adapted screening for ESBL-E carriage is performed, but limited data exist on the optimal screening strategy. Currently, laboratory detection of ESBL-E in fecal specimens mostly relies on culture methods using selective agar medium. This method has a modest sensitivity (3). Preenrichment of swabs is used by many laboratories for methicillin-resistant Staphylococcus aureus (MRSA) screening and has demonstrated significantly improved sensitivity (4). To our knowledge, the impact of preenrichment on ESBL-E detection in stool samples has not been systematically investigated. In our study, we compared three different selective and nonselective broths for the detection of ESBL-E in clinical stool samples to the standard procedure-direct plating on ESBL agar.(This study was presented in part as a poster at the 25th European Congress of Clinical Microbiology and Infectious Diseases, Copenhagen, Denmark, 25 to 28 April 2015.) From September 2013 to March 2014, 496 stool samples submitted for ESBL-E screening from 343 patients at the University Hospital Cologne, a 1,400-bed tertiary care facility, were included in the study. All samples were stored at 4°C and were processed within 24 h of receipt. A pea-sized portion of solid stool or 250 l of liquid stool was suspended in 1 ml 0.9% NaCl to reach a standardized inoculum. One hundred microliters of the stool suspension was inoculated directly onto chromID ESBL agar (bioMérieux, Marcy l'Etoile, France) and into 5 ml of nonselective tryptic soy broth (TSB), MacConkey (MC) broth (Roth, Germany, Karlsruhe) selecting only Gram-negative rods, and MacConkey broth supplemented with cefuroxime (32 mg/liter) and vancomycin (64 mg/liter) (MC-CV) additionally inhibiting the nonresistant Gram-negative flora and further inhibiting Grampositive bacteria. Each medium was stored and processed according to the manufacturers' instructions.Agar plates and broths were incubated at 36°C Ϯ 1°C in ambient air for 18 h to 24 h. Using calibrated inoculation loops, 10 l of the respective enrichment broth was inoculated onto ESBL agar, which was incubated at 36°C Ϯ 1°C in ambient air for another 18 h to 24 h. Plates were interpreted according to the manufacturer's instructions. Uncolored colonies growing on ...

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Rectal Carriage of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae in Hospitalized Patients: Selective Preenrichment Increases Yield of Screening

Cited by 55 publications

References 21 publications

Antimicrobial resistant and extended‐spectrum β‐lactamase producing Escherichia coli in common wild bird species in Switzerland

Antimicrobial resistant and extended‐spectrum β‐lactamase producing Escherichia coli in common wild bird species in Switzerland

Whole-Genome Multilocus Sequence Typing of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae

Use of an Enrichment Broth Improves Detection of Extended-Spectrum-Beta-Lactamase-Producing Enterobacteriaceae in Clinical Stool Samples

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