Complete Genome Sequence of
            <i>Escherichia coli</i>
            81009, a Representative of the Sequence Type 131 C1-M27 Clade with a Multidrug-Resistant Phenotype

Mutti, Michele; Sonnevend, Ágnes; Pál, Tibor; Junttila, Sini; Ekker, Heinz; Gálik, Bence; Gyenesei, Attila; Nagy, Gábor; Nagy, Eszter; Szijártó, Valéria

doi:10.1128/genomea.00056-18

Cited by 8 publications

(5 citation statements)

References 12 publications

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“…E. coli strains 81009 (14, 15) and MG1655 (16) were grown in LB or on Trypticase soy agar (TSA) plates (bioMérieux). To prepare overnight cultures, a single colony was inoculated in LB and incubated at 37 °C with agitation at 200 RPM overnight.…”

Section: Methodsmentioning

confidence: 99%

A New Tool for Complement Research: In vitro Reconstituted Human Classical Complement Pathway

et al. 2018

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The complement, as part of the innate immune system, represents the first line of defense against Gram-negative bacteria invading the bloodstream. The complement system is a zymogen cascade that ultimately assemble into the so-called membrane attack complex (MAC), which lyses Gram-negative bacteria upon insertion into the outer membrane. Traditionally, serum has been used as complement source, for example to study the bactericidal activity of monoclonal antibodies or antibodies raised upon vaccination. Due to the significant donor to donor variability, as well as susceptibility of complement factors to handling and storage conditions, assay reproducibility using human serum is low. Moreover, the presence of pre-existing antibodies and antimicrobial compounds are confounding factors. To remove antibodies from human serum, we applied κ/λ-light chain specific affinity chromatography, however the method severely reduced the complement activity due to the depletion of complement components. Therefore, we attempted to reconstitute human complement—namely the alternative (rAP) and the classical (rCP) pathways—from purified complement factors. We found that adding C1-inhibitor to the mixture was essential to maintain a stable and functional C1 and thus to generate an active rCP. We further confirmed the functionality of the rCP by testing the complement-dependent bactericidal activity of a human monoclonal antibody, A1124 against an E. coli clinical isolate belonging to the ST131 clonal complex, and that of a polyclonal IVIg against a laboratory E. coli strain (MG1655) not expressing LPS O-antigen and capsule. Although the alternative pathway did not have any bactericidal activity by itself, it enhanced MAC deposition induced by rCP and increased the overall bactericidal activity against the ST131 E. coli strain. In conclusion, we report for the first time the successful in vitro reconstitution of the classical pathway of the human complement to establish a serum-free, complement dependent bactericidal assay. This system offers high level of standardization and could support the study of the complement in different research fields.

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Section: Methodsmentioning

confidence: 99%

A New Tool for Complement Research: In vitro Reconstituted Human Classical Complement Pathway

et al. 2018

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“…Cluster type (CT) for E. coli was defined as isolates attributed to the same clone with ≤ 10 differing alleles (CT threshold) within a group. The ST131 isolates were compared to three international ST131 E. coli genomes (C1-M27: EC81009 [16] and H105 [17], C2/H30Rx: JJ2434 [18]) obtained from GenBank, to reveal the relatedness to the subclades of the ST131 clone. The replicon types were retrieved using PlasmidFinder2.1 and pMLST-2.0 [19] online tools at the Center for Genomic Epidemiology (CGE, http:// genom icepi demio logy.…”

Section: Molecular Characterizationmentioning

confidence: 99%

Population snapshot of the extended-spectrum β-lactamase-producing Escherichia coli invasive strains isolated from a Hungarian hospital

Tóth

Kamotsay³

et al. 2022

Ann Clin Microbiol Antimicrob

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Background This study was carried out to determine the prevalence and the genetic background of extended-spectrum β-lactamase-producing Escherichia coli invasive isolates obtained from a tertiary-care hospital in Budapest, Hungary. Methods Between October–November 2018, all invasive ESBL-producing E. coli isolates were collected from Central Hospital of Southern Pest. The antimicrobial susceptibility testing was performed according to the EUCAST guidelines. The possible clonal relationships were investigated by core genome (cg)MLST (SeqSphere +) using whole-genome sequencing (WGS) data of isolates obtained from Illumina 251-bp paired-end sequencing. From WGS data acquired antimicrobial resistance genes, virulence genes and replicon types were retrieved using ResFinder3.1, PlasmidFinder2.1, pMLST-2.0, VirulenceFinder2.0 and Virulence Factors Database online tools. Results Overall, six E. coli isolates proved to be resistant to third-generation cephalosporins and ESBL-producers in the study period. Full genome sequence analysis showed that five E. coli isolates belonged to the ST131 clone: two to C1-M27 subclade with blaCTX-M-27 and three to C2/H30Rx subclade with blaCTX-M-15. One isolate belonged to ST1193 with blaCTX-M-27. According to cgMLST, all C2/H30Rx isolates formed a cluster (≤ 6 allele differences), while the blaCTX-M-27-producing C1-M27 isolates differed at least 35 alleles from each other. Both C2/H30Rx and C1-M27 ST131 isolates harbored similar antimicrobial resistance gene sets. However, only C2/H30Rx isolates had the qnrB and aac(3)-IIa. The isolates carried similar extraintestinal virulence gene set but differed in some genes encoding siderophores, protectins and toxins. Moreover, only one C2/H30Rx isolate carried salmochelin siderophore system and showed virotype B. All isolates showed resistance against ceftriaxone, cefotaxime, and ciprofloxacin, and the C2/H30Rx isolates were also resistant to gentamicin, tobramycin, and ceftazidime. Conclusions Out of six ESBL-producing E. coli, five belonged to the ST131 clone. This study indicates, that the C2/H30Rx and C1-M27 subclades of the ST131 appear to be the dominant clones collected in a Hungarian hospital.

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“…Cluster type (CT) for E. coli was de ned as isolates attributed to the same clone with ≤ 10 differing alleles (CT threshold) within a group. The ST131 isolates were compared to three international ST131 E. coli genomes (C1-M27: EC81009 [15] and H105 [16], C2/H30Rx: JJ2434 [17]) obtained from GenBank, to reveal the relatedness to the subclades of the ST131 clone. The replicon types were retrieved using PlasmidFinder2.1 and pMLST-2.0 [18] online tools at the Center for Genomic Epidemiology (CGE, http://genomicepidemiology.org/).…”

Section: Molecular Characterizationmentioning

confidence: 99%

Population Snapshot of the Extended-spectrum β-lactamase-producing Escherichia Coli Invasive Strains Isolated From a Hungarian Hospital

Tóth

Kamotsay³

et al. 2021

Preprint

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Background:This study was carried out to determine the prevalence and the genetic background of extended-spectrum β-lactamase-producing Escherichia coli invasive isolates obtained from a tertiary-care hospital in Budapest, Hungary. MethodsBetween October-November 2018, all invasive ESBL-producing E. coli isolates were collected from Central Hospital of Southern Pest. The antimicrobial susceptibility testing was performed according to the EUCAST guidelines. The possible clonal relationships were investigated by core genome (cg)MLST (SeqSphere+) using whole-genome sequencing (WGS) data of isolates obtained from Illumina 251-bp paired-end sequencing. From WGS data acquired antimicrobial resistance genes and replicon types were retrieved using ResFinder3.1, PlasmidFinder2.1, and pMLST-2.0 online tools.ResultsOverall, 25 E. coli isolates were detected and six proved to be resistant to third-generation cephalosporins. Full genome sequence analysis showed that five E. coli isolates belonged to the ST131 clone: two to C1-M27 subclade with blaCTX-M-27 and three to C2/H30Rx subclade with blaCTX-M-15. One isolate belonged to ST1193 with blaCTX-M-27. According to cgMLST, all C2/H30Rx isolates formed a cluster (≤6 allele differences), while the blaCTX-M-27-producing C1-M27 isolates differed at least 35 alleles from each other. Both C2/H30Rx and C1-M27 ST131 isolates harbored similar antimicrobial resistance gene sets. However, only C2/H30Rx isolates had the qnrB and aac(3)-IIa. All isolates showed resistance against ceftriaxone, cefotaxime, and ciprofloxacin, and the C2/H30Rx isolates were also resistant to gentamicin, tobramycin, and ceftazidime.ConclusionsOut of six third-generation cephalosporins-resistant, invasive E. coli, five belonged to the S131clone. This study indicates, that the C2/H30Rx and C1-M27 subclades of the ST131 appear to be the dominant clones collected in a Hungarian hospital.

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Complete Genome Sequence of Escherichia coli 81009, a Representative of the Sequence Type 131 C1-M27 Clade with a Multidrug-Resistant Phenotype

Cited by 8 publications

References 12 publications

A New Tool for Complement Research: In vitro Reconstituted Human Classical Complement Pathway

A New Tool for Complement Research: In vitro Reconstituted Human Classical Complement Pathway

Population snapshot of the extended-spectrum β-lactamase-producing Escherichia coli invasive strains isolated from a Hungarian hospital

Population Snapshot of the Extended-spectrum β-lactamase-producing Escherichia Coli Invasive Strains Isolated From a Hungarian Hospital

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