Persistence of a Distinct
            <i>Corynebacterium diphtheriae</i>
            Clonal Group within Two Communities in the United States and Canada Where Diphtheria Is Endemic

Marston, Chung K.; Jamieson, Frances; Cahoon, Fred; Lesiak, Gail; Golaz, Anne; Reeves, Mike; Popović, Tanja

doi:10.1128/jcm.39.4.1586-1590.2001

Cited by 36 publications

(10 citation statements)

References 15 publications

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“…It has been suggested that persistent foci of diphtheria in Russia could be a possible source of the epidemic strains since Russia was never totally free of reported cases of diphtheria (39). Reports of persistent endemic foci in the United States (24,31) and Canada (24) suggest that the circulation of toxigenic strains of C. diphtheriae can occur for prolonged periods even in the absence of recognized clinical cases, at least in certain communities. Finally, we suggest that such permanent isolation of C. diphtheriae strains of ribotypes Sankt-Peterburg and Rossija in Russia reflects a stable endemicity of this clonal group within this geographic area.…”

Section: Discussionmentioning

confidence: 99%

Efficient Discrimination within aCorynebacterium diphtheriaeEpidemic Clonal Group by a Novel Macroarray-Based Method

et al. 2005

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The diphtheria epidemic in Russia and neighboring countries in the 1990s (140,000 cases, 4,000 deaths in 1991 to 1996 [39]) stimulated research activities on Corynebacterium diphtheriae, a causative agent of the disease. A number of the typing methods available at that time (multilocus enzyme electrophoresis [MLE], pulsed-field gel electrophoresis [PFGE], ribotyping, and randomly amplified polymorphic DNA [RAPD] analysis) and newer methods (amplified fragment length polymorphism analysis) were applied for interstrain differentiation of the pathogen (6,7,8,20,22,23,28,29,33,36,40). These methods allowed the identification of a clonal group of closely related strains responsible for the epidemic in Russia and all other countries of the former Soviet Union and to trace strains exported into other countries (6,22,23,32,33). These strains were indistinguishable by PFGE, RAPD analysis, and amplified fragment length polymorphism analysis and very similar by ribotyping (there were two principal profiles, "Rossija" and "Sankt-Peterburg," which differed by one band [6, 14, 33]). Minor rare variants were identified by RAPD and ribotyping techniques (22), and a total of 27 types similar by Ͼ80% were identified by MLE typing of all strains of this clonal group studied to date (32, 33). However, MLE, PFGE, and ribotyping are time-consuming and rather cumbersome methods, while RAPD analysis lacks interlaboratory reproducibility and hence exchangeability of results. To identify and rapidly monitor subtle changes in the genome structure at an infraclonal level during and between epidemics, fast, simple, portable, and discriminatory molecular typing methods of C. diphtheriae are still needed.Repetitive genome sequences present important sources of intraspecies variation. A new family of such loci (clustered, regularly interspaced, short palindromic repeats [CRISPR]) has recently been identified by in silico analysis of many bacterial species (19). This family is characterized by direct repeats (DR) varying in size from 21 to 37 bp, interspaced by similarly sized nonrepetitive sequences (variable spacers). DR and adjacent variable spacers form direct variant repeats (DVR) (20). The DNA reverse-hybridization method was developed to study variation in the Mycobacterium tuberculosis DR locus (the presence or absence of 43 different spacers) by using the macroarray format; this method was named "spoligotyping" (spacer oligonucleotide typing [20]) and has been widely used for epidemiological and phylogenetic purposes (12,20,35).In 2003, a complete genome sequence of the C. diphtheriae epidemic strain of biotype gravis ribotype Sankt-Peterburg was published (4). This publication made possible a more thorough, precise, and comprehensive search of candidate polymorphic loci for the development of new typing methods for this pathogen. In the present study, we identified in silico a large DR region in the genome of C. diphtheriae and developed a reverse-hybridization macroarray-based method to study its polymorphism. Using this method, we e...

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Section: Discussionmentioning

confidence: 99%

Efficient Discrimination within aCorynebacterium diphtheriaeEpidemic Clonal Group by a Novel Macroarray-Based Method

et al. 2005

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“…We performed PCR and Western blotting (WB) to examine toxigenicity as well as the presence of major pilin genes and their expression in 42 clinical isolates collected between 1979 and 2000 from different parts of the world (4,16,(20)(21)(22)(23). For PCR analysis, primers targeting the toxin gene (tox) and the major pilin subunits of each pilus type (spaA, spaD, and spaH) were designed based on the sequence of the type strain NCTC13129 (8).…”

Section: Resultsmentioning

confidence: 99%

“…To obtain a better insight into the roles of corynebacterial pili in pathogenesis, we conducted a comparative functional genomic analysis using 42 C. diphtheriae clinical isolates gathered mostly from Russia, Canada, and the United States (20)(21)(22)(23). The pilus gene pool was assessed by PCR amplification using primers for spa genes of NCTC13129, and pilus expression was monitored by immunoblotting as well as immunoelectron microscopy (IEM) using antibodies against respective Spa pilins.…”

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confidence: 99%

Pilus Gene Pool Variation and the Virulence of Corynebacterium diphtheriae Clinical Isolates during Infection of a Nematode

et al. 2013

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c Toxigenic Corynebacterium diphtheriae strains cause diphtheria in humans. The toxigenic C. diphtheriae isolate NCTC13129 produces three distinct heterotrimeric pili that contain SpaA, SpaD, and SpaH, making up the shaft structure. The SpaA pili are known to mediate bacterial adherence to pharyngeal epithelial cells. However, to date little is known about the expression of different pili in various clinical isolates and their importance in bacterial pathogenesis. Here, we characterized a large collection of C. diphtheriae clinical isolates for their pilin gene pool by PCR and for the expression of the respective pilins by immunoblotting with antibodies against Spa pilins. Consistent with the role of a virulence factor, the SpaA-type pili were found to be prevalent among the isolates, and most significantly, corynebacterial adherence to pharyngeal epithelial cells was strictly correlated with isolates that were positive for the SpaA pili. By comparison, the isolates were heterogeneous for the presence of SpaD-and SpaHtype pili. Importantly, using Caenorhabditis elegans as a model host for infection, we show here that strain NCTC13129 rapidly killed the nematodes, the phenotype similar to isolates that were positive for toxin and all pilus types. In contrast, isogenic mutants of NCTC13129 lacking SpaA-type pili or devoid of toxin and SpaA pili exhibited delayed killing of nematodes with similar kinetics. Consistently, nontoxigenic or toxigenic isolates that lack one, two, or all three pilus types were also attenuated in virulence. This work signifies the important role of pili in corynebacterial pathogenesis and provides a simple host model to identify additional virulence factors.

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“…In 2001, Marston et al (12) reported on the circulation of toxigenic endemic strains of C. diphtheriae within two communities in the United States and Canada for at least 25 years. The reason why certain strains of C. diphtheriae cause outbreaks and others remain endemic is a subject for speculation.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Four Molecular Typing Methods for Characterization of Corynebacterium diphtheriae and Determination of Transcontinental Spread of C. diphtheriae Based on BstEII rRNA Gene Profiles

Zoysa¹,

Hawkey

Charlett

et al. 2008

J Clin Microbiol

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The diphtheria epidemic in the Russian Federation in the 1990s made diphtheria a focus of global concern once again. The development of rapid and reproducible typing methods for the molecular characterization of Corynebacterium diphtheriae has become a priority in order to be able to monitor the spread of this important pathogen on a global scale. We report on a comparison of four molecular typing methods (ribotyping, pulsed-field gel electrophoresis [PFGE], random amplification of polymorphic DNA [RAPD], and amplified fragment length polymorphism [AFLP]) for the characterization of C. diphtheriae strains. Initially, 755 isolates originating from 26 countries were analyzed by ribotyping. One strain of each ribotype was then randomly chosen and characterized by PFGE, RAPD, and AFLP. In order to ascertain whether the Eastern European epidemic ribotype could be further discriminated, 10 strains of ribotype D1 (the epidemic ribotype) from different geographical regions were randomly chosen and subjected to analysis by PFGE, RAPD, and AFLP. The results revealed that ribotyping is highly discriminatory and reproducible and is currently the method of choice for typing C. diphtheriae. PFGE and AFLP were less discriminatory than ribotyping and RAPD. An assessment of the transcontinental spread of the organism showed that several genotypes of C. diphtheriae circulated on different continents of the world and that each outbreak was caused by a distinct clone. The ribotypes seen in Europe appeared to be distinct from those seen elsewhere, and certain ribotypes appeared to be unique to particular countries.

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Persistence of a Distinct Corynebacterium diphtheriae Clonal Group within Two Communities in the United States and Canada Where Diphtheria Is Endemic

Cited by 36 publications

References 15 publications

Efficient Discrimination within aCorynebacterium diphtheriaeEpidemic Clonal Group by a Novel Macroarray-Based Method

Efficient Discrimination within aCorynebacterium diphtheriaeEpidemic Clonal Group by a Novel Macroarray-Based Method

Pilus Gene Pool Variation and the Virulence of Corynebacterium diphtheriae Clinical Isolates during Infection of a Nematode

Comparison of Four Molecular Typing Methods for Characterization of Corynebacterium diphtheriae and Determination of Transcontinental Spread of C. diphtheriae Based on BstEII rRNA Gene Profiles

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