Investigation of an outbreak of antibiotic-associated colitis by various typing methods

Wüst, J; Sullivan, Nadine M.; Hardegger, U; Wilkins, Tracy D.

doi:10.1128/jcm.16.6.1096-1101.1982

Cited by 137 publications

(37 citation statements)

References 38 publications

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“…spread of C. difficile, particularly when the organism is isolated from patients and their hospital environment (5, 6, 9), the need to establish a C. difficile typing method to elucidate the epidemiology of this organism remains. Several workers have initiated various schemes to investigate outbreaks of diarrhea associated with antibiotic therapy and chemotherapy (14, 19), and others have described various methods for typing the organism (2, 13, 16,17). In this paper we report a typing method based on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins.…”

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confidence: 99%

Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins

Tabaqchali¹,

O'Farrell²,

Holland³

et al. 1986

J Clin Microbiol

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A typing method for Clostridium difficile based on the incorporation of [35S]methionine into cellular proteins, their separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their visualization by autoradiography is described. On analysis of the radiolabeled-protein profiles, nine distinct groups were observed (A to E and W to Z). The method, which is simple, reproducible, and readily expandable, has been applied in epidemiological studies to demonstrate cross-infection and hospital acquisition of C. ditficile. The epidemiology of Clostridiium difficile diarrheal disease has not yet been fully clarified. Although C. difficile is known to be the primary cause of pseudomembranous colitis and antibiotic-associated colitis (AAC) and is associated with diarrhea after antibiotic therapy and chemotherapy (1, 8, 10, 11), this organism has also been isolated from the feces of healthy adults (0 to 11%) (1, 3, 12), from vaginal specimens of healthy mothers (18%) (18), and from asymptomatic infants and neonates (40 to 75%) (4, 7, 12, 15, 18). Although reports of clustering of cases and outbreaks of pseudomembranous colitis and AAC are suggestive of the nosocomial

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confidence: 99%

Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins

Tabaqchali¹,

O'Farrell²,

Holland³

et al. 1986

J Clin Microbiol

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“…The 630 strain was isolated in 1982, so passaging within and between labs could have led to accumulation of mutations that reduced the frequency of flagellar phase variation in 630. We thus asked whether the lack of (or very low frequency of) flagellar switch inversion in 630 is unique to this strain and its derivatives, or a broader attribute of currently circulating ribotype 012 C. difficile isolates (18). To test this, we evaluated flagellar phase variation in six riboype 012 isolates: SS235, a community environmental isolate; SE838, a hospital environmental isolate; and MAM30, MT4768, MT5065, and MT5066, isolates from patients with CDI (kindly gifted by Dr. Kevin Garey).…”

Section: Resultsmentioning

confidence: 99%

“…For C. difficile strains R20291, a ribotype 027 strain, and ATCC 43598, a ribotype 017 strain, both the flg ON and OFF orientations were apparent under multiple conditions tested. However, only the flg ON orientation was detectable in 630, a ribotype 012 strain originally isolated from a patient with C. difficile infection (18). However, recent work from Collery et al suggests the flagellar switch is capable of inversion in 630 (19), suggesting this strain is phase-locked.…”

Section: Introductionmentioning

confidence: 99%

Characterization of flagellar and toxin phase variation inClostridium difficileribotype 012 isolates

Anjuwon-Foster

Maldonado-Vazquez

Tamayo

2018

Preprint

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20Clostridium difficile causes diarrheal diseases mediated in part by the secreted 21 toxins TcdA and TcdB. C. difficile produces flagella that also contribute to motility and 22 bacterial adherence to intestinal cells during infection. Flagellum and toxin gene 23 expression are linked via the flagellar alternative sigma factor, SigD. Recently, we 24 identified a "flagellar switch" upstream of the early flagellar biosynthesis operon that 25 mediates phase variation of both flagellum and toxin production in C. difficile strain 26 R20291. However, we were unable to detect flagellar switch inversion in C. difficile 27 strain 630, a ribotype 012 strain commonly used in research labs, suggesting the strain 28 is phase-locked. To determine whether a "phase locked" flagellar switch is limited to 630 29 or present more broadly in ribotype 012 strains, we assessed the frequency and 30 phenotypic outcomes of flagellar switch inversion in multiple C. difficile ribotype 012 31isolates. The laboratory-adapted strain JIR8094, a derivative of strain 630, and six 32 clinical and environmental isolates were all found to be phase OFF, non-motile, and 33 attenuated for toxin production. We isolated low frequency motile derivatives of JIR8094 34 with partial recovery of motility and toxin production, and found that additional changes 35 in JIR8094 impact these processes. The clinical and environmental isolates varied 36 considerably in the frequency by which flagellar phase ON derivatives arose, and these 37 derivatives showed fully restored motility and toxin production. Taken together, these 38 results demonstrate heterogeneity in flagellar and toxin phase variation among C.

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“…TcdB2 has enhanced ability to enter host cells, is more cytotoxic, and exhibits wider tissue tropism (44, 45). In contrast to RT027 strains, the PaLoc of RT106 strains is 100% identical to 630 (53-56); thus, these strains encode full-length TcdC and express the TcdB1 toxin variant. Both RT027 and RT106 isolates produce variable amounts of TcdA/TcdB.…”

Section: Discussionmentioning

confidence: 99%

Phylogenomic analysis ofClostridiodes difficileribotype 106 strains reveals novel genetic islands and emergent phenotypes

Roxas

Claus-Walker

et al. 2020

Preprint

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27Background: Clostridioides difficile RT106 has emerged as a dominant molecular type 28 in the USA in recent years, but the underlying factors contributing to its predominance 29 remain undefined. As part of our ongoing C. difficile infection (CDI) surveillance in 30 Arizona, we monitored RT106 frequency and characterized the genomic and phenotypic 31 properties of the recovered isolates. 32Results: From 2015-2018, RT106 was the second-most prevalent molecular type 33 isolated from CDI patients in our surveillance. A representative RT106 strain displayed 34 robust virulence and 100% lethality in the hamster model of acute CDI. We identified a 35 unique 46 KB genomic island (GI1) in all RT106 strains, including those in public 36 databases. GI1 was not found in its entirety in any other C. difficile clade, or indeed in any 37 other microbial genome; however, smaller segments were detected in select 38 Enterococcus faecium strains. Molecular clock analyses suggest that GI1 was 39 horizontally acquired and sequentially assembled over time. Consistent with the presence 40 of genes encoding homologs of VanZ and a SrtB-anchored collagen-binding adhesin in 41 GI1, all tested RT106 strains had increased teicoplanin resistance and a majority 42 displayed collagen-dependent biofilm formation. Two additional genomic islands (GI2 and 43 GI3) were also present in a subset of RT106 strains. All three islands have features of 44 mobile genetic elements and encode several putative virulence factors. 45 Conclusions: Consistent with the known genetic plasticity of C. difficile, strains belonging 46 to the RT106 clade harbor unique genetic islands. Correspondingly, emergent phenotypic 47 properties may contribute to the relatively rapid shifts of strain distribution in patient 48 populations. 49 Keywords: Clostridioides difficile, RT106, teicoplanin, VanZ, SrtB-anchored collagen-50 binding adhesin, genomic island 51 52 BACKGROUND 53The Gram-positive and spore-forming anaerobic bacterium Clostridioides difficile 54 (formerly named Clostridium difficile) is a leading cause of antibiotic-associated diarrhea 55 that may be self-limiting, or progress to severe and fulminant (pseudomembranous) colitis 56 or toxic megacolon (1-4). There has been an increased incidence of C. difficile infection 57 (CDI) over the past two decades (5-8) and, in the USA, this coincides with the emergence 58 and spread of ribotype 027 strains [also called RT027 or BI or NAP1 based on the 59 phylogenetic test (9, 10)]. While RT027 remains the most prevalent healthcare-associated 60 C. difficile ribotype, its frequency has been steadily declining (11). Multiple surveillance 61 studies indicate a changing trend in the C. difficile ribotype frequency distribution, 62 particularly the emergence of RT106 (also called Group "DH" or "NAP11") in regions 63 where it was previously rarely found. In 2008, RT106 was second to RT027 as the most 64 dominant ribotype in England, and was also identified in neighboring European countries 65 including . Howeve...

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Investigation of an outbreak of antibiotic-associated colitis by various typing methods

Cited by 137 publications

References 38 publications

Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins

Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of [35S]methionine-labeled proteins

Characterization of flagellar and toxin phase variation inClostridium difficileribotype 012 isolates

Phylogenomic analysis ofClostridiodes difficileribotype 106 strains reveals novel genetic islands and emergent phenotypes

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