New Flagellin-Specifying Genes in Some
            <i>Escherichia coli</i>
            Strains

Ratiner, Yuli A.

doi:10.1128/jb.180.4.979-984.1998

Cited by 29 publications

(12 citation statements)

References 32 publications

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“…4). Others were randomly assembled into eight reaction 5 kits. In multiple PCR assays, the same results as those for simplex PCR assays were obtained from all reference strains.…”

Section: Resultsmentioning

confidence: 99%

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Escherichia coli H-Genotyping PCR: a Complete and Practical Platform for Molecular H Typing

et al. 2018

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In , more than 180 O groups and 53 H types have been recognized. The O:H serotyping of strains is an effective method for identifying strains with pathogenic potential and classifying them into clonal groups. In particular, the serotyping of Shiga toxin-producing (STEC) strains provides valuable information to evaluate the routes, sources, and prevalence of agents in outbreak investigations and surveillance. Here, we present a complete and practical PCR-based H-typing system, H-genotyping PCR, consisting of 10 multiplex PCR kits with 51 single PCR primer pairs. Primers were designed based on a detailed comparative analysis of sequences from all H-antigen (flagellin)-encoding genes, and its homologs. The specificity of this system was confirmed by using all H type reference strains. Additionally, 362 serotyped wild strains were also used to evaluate its practicality. All 277 H-type-identified isolates gave PCR products that corresponded to the results of serological H typing. Moreover, 76 nonmotile and nine untypeable strains could be successfully subtyped into any H type by the PCR system. The H-genotyping PCR developed here allows broader, rapid, and low-cost subtyping of H types and will assist epidemiological studies as well as surveillance of pathogenic .

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“…4). Others were randomly assembled into eight reaction 5 kits. In multiple PCR assays, the same results as those for simplex PCR assays were obtained from all reference strains.…”

Section: Resultsmentioning

confidence: 99%

“…In E. coli, flagellin is encoded by the fliC gene on the chromosomal locus or its homologues (non-fliC flagellin-coding genes) at loci other than fliC, such as flkA, fllA, and flmA (4)(5)(6)(7)(8). When a mobile element carrying non-fliC and its attendant gene is inserted in the chromosome, these genes may mediate reversible flagellar phase variation.…”

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confidence: 99%

Escherichia coli H-Genotyping PCR: a Complete and Practical Platform for Molecular H Typing

et al. 2018

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“…In total, 139 (74%) isolates were motile in soft agar, of which 25 (18%) isolates showed no amplification of the fliC gene. This could be due to sequence variations in fliC among different E. coli isolates (40) and/or the expression of another or new flagellin genes (41). Although Pratt and Kolter found that motility, and not chemotaxis, is required for E. coli to form a biofilm (21), it is also possible for nonmotile strains to adhere to surfaces and develop a biofilm (42).…”

Section: Discussionmentioning

confidence: 99%

Genotypic and Phenotypic Characteristics Associated with Biofilm Formation by Human Clinical Escherichia coli Isolates of Different Pathotypes

Schiebel

Böhm

Nitschke

et al. 2017

Appl Environ Microbiol

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Bacterial biofilm formation is a widespread phenomenon and a complex process requiring a set of genes facilitating the initial adhesion, maturation, and production of the extracellular polymeric matrix and subsequent dispersal of bacteria. Most studies on biofilm formation have investigated nonpathogenic K-12 strains. Due to the extensive focus on laboratory strains in most studies, there is poor information regarding biofilm formation by pathogenic isolates. In this study, we genotypically and phenotypically characterized 187 human clinical isolates representing various pathotypes (e.g., uropathogenic, enteropathogenic, and enteroaggregative ). We investigated the presence of biofilm-associated genes ("genotype") and phenotypically analyzed the isolates for motility and curli and cellulose production ("phenotype"). We developed a new screening method to examine the biofilm formation ability. In summary, we found a high prevalence of biofilm-associated genes. However, we could not detect a biofilm-associated gene or specific phenotype correlating with the biofilm formation ability. In contrast, we did identify an association of increased biofilm formation with a specific pathotype. Enteroaggregative (EAEC) was found to exhibit the highest capacity for biofilm formation. Using our image-based technology for the screening of biofilm formation, we demonstrated the characteristic biofilm formation pattern of EAEC, consisting of thick bacterial aggregates. In summary, our results highlight the fact that biofilm-promoting factors shown to be critical for biofilm formation in nonpathogenic strains do not reflect their impact in clinical isolates and that the ability of biofilm formation is a defined characteristic of EAEC. Bacterial biofilms are ubiquitous and consist of sessile bacterial cells surrounded by a self-produced extracellular polymeric matrix. They cause chronic and device-related infections due to their high resistance to antibiotics and the host immune system. In nonpathogenic , cell surface components playing a pivotal role in biofilm formation are well known. In contrast, there is poor information for their role in biofilm formation of pathogenic isolates. Our study provides insights into the correlation of biofilm-associated genes or specific phenotypes with the biofilm formation ability of commensal and pathogenic Additionally, we describe a newly developed method enabling qualitative biofilm analysis by automated image analysis, which is beneficial for high-throughput screenings. Our results help to establish a better understanding of biofilm formation.

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“…PCR detection of fliC alleles has been used for molecular H-typing for some time (Wang et al ., 2003). However, some E. coli isolates have an alternative flagellar phase, due to the presence of an additional flagellin gene (flnaA, fllA, fmlA or flkA) , similar to those found in Salmonella species (Feng et al ., 2008; Ratiner, 1998; Ratiner et al ., 2010; Tominaga, 2004; Tominaga & Kutsukake, 2007).…”

Section: Introductionmentioning

confidence: 99%

EcOH:In silicoserotyping ofE. colifrom short read data

Ingle

Valcanis

Kuzevski

et al. 2015

Preprint

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22"! 23"The lipopolysaccharide (O) and flagellar (H) surface antigens of Escherichia coli are targets 24"for serotyping that have traditionally been used to identify pathogenic lineages of E. coli. As 25"serotyping has several limitations, public health reference laboratories are increasingly 26"moving towards whole genome sequencing (WGS) for the rapid characterisation of bacterial 27"isolates. Here we present a method to rapidly and accurately serotype E. coli isolates from 28"raw, short read sequence data, leveraging the known genetic basis for the biosynthesis of O- 29"and H-antigens. Our approach bypasses the need for de novo genome assembly by directly 30"screening WGS reads against a curated database of alleles linked to known E. coli O-groups 31"and H-types (the EcOH database) using the software package SRST2. We validated our 32"approach by comparing in silico results with those obtained via serological phenotyping of 33"197 enteropathogenic (EPEC) isolates. We also demonstrated the utility of our method to 34"characterise enterotoxigenic E. coli (ETEC) and the uropathogenic E. coli (UPEC) epidemic 35"clone ST131, and for in silico serotyping of foodborne outbreak-related isolates in the public 36"GenomeTrakr database.

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New Flagellin-Specifying Genes in Some Escherichia coli Strains

Cited by 29 publications

References 32 publications

Escherichia coli H-Genotyping PCR: a Complete and Practical Platform for Molecular H Typing

Escherichia coli H-Genotyping PCR: a Complete and Practical Platform for Molecular H Typing

Genotypic and Phenotypic Characteristics Associated with Biofilm Formation by Human Clinical Escherichia coli Isolates of Different Pathotypes

EcOH:In silicoserotyping ofE. colifrom short read data

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