13,043 research outputs found

    Epidemiology of Shigella-Associated diarrhea in Gorgan, north of Iran

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    Objective : Shigella is an important etiological agent for diarrhea and especially dysentery. Shigellosis is an intestinal infection that is a major public health problem in many developing countries. The aim of this study was to evaluate the prevalence of Shigella and its various species in diarrheal samples in Gorgan located in the north of Iran. Materials and Methods: Between January-December 2005, the epidemiology of Shigella- associated diarrhea was studied among 634 patients in Gorgan. The diarrheal samples accompanied with a questionnaire, which contained the demographic and main symptoms of the patients, were transported to the laboratory and inoculated in different culture media. Colonies suspected to be of Shigella were detected using differential biochemical tests and subsequently, the serotype of Shigella was defined using antisera. Results : Shigella was isolated from 56/634 diarrheal samples) (8.8%) of which S. sonnei was the predominant species (55%). Occurrence of Schigella was highest in the 2-5 years′ age group (70.9%) and highest in summer (73.2%) with the most frequent clinical manifestation being abdominal pain (67.8%). The prevalence of Shigella in males and females was 8 and 9.8% respectively, but this difference was not statistically significant. Conclusion: It has been shown that Shigella sonnei is the most common Shigella serogroup among 2-5 year-old children in Gorgan. It is therefore suggested that hygienic training be given to childcare attendants and the children themselves

    A Pentaplex PCR Assay for the Detection and Differentiation of Shigella Species

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    The magnitude of shigellosis in developing countries is largely unknown because an affordable detection method is not available. Current laboratory diagnosis of Shigella spp. is laborious and time consuming and has low sensitivity. Hence, in the present study, a molecular-based diagnostic assay which amplifies simultaneously four specific genes to identify invC for Shigella genus, rfc for S. flexneri, wbgZ for S. sonnei, andrfpB for S. dysenteriae, aswell as one internal control (ompA) gene, was developed in a single reaction to detect and differentiate Shigella spp. Validation with 120 Shigella strains and 37 non-Shigella strains yielded 100% specificity. The sensitivity of the PCR was 100 pg of genomic DNA, 5.4 × 104 CFU/ml, or approximately 120 CFU per reaction mixture of bacteria. The sensitivity of the pentaplex PCR assay was further improved following preincubation of the stool samples in Gram-negative broth. A preliminary study with 30 diarrhoeal specimens resulted in no cross-reaction with other non-Shigella strains tested. We conclude that the developed pentaplex PCR assay is robust and can provide information about the four target genes that are essential for the identification of the Shigella genus and the three Shigella species responsible for the majority of shigellosis cases

    The Multifaceted Activity of the VirF Regulatory Protein in the Shigella Lifestyle

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    Shigella is a highly adapted human pathogen, mainly found in the developing world and causing a severe enteric syndrome. The highly sophisticated infectious strategy of Shigella banks on the capacity to invade the intestinal epithelial barrier and cause its inflammatory destruction. The cellular pathogenesis and clinical presentation of shigellosis are the sum of the complex action of a large number of bacterial virulence factors mainly located on a large virulence plasmid (pINV). The expression of pINV genes is controlled by multiple environmental stimuli through a regulatory cascade involving proteins and sRNAs encoded by both the pINV and the chromosome. The primary regulator of the virulence phenotype is VirF, a DNA-binding protein belonging to the AraC family of transcriptional regulators. The virF gene, located on the pINV, is expressed only within the host, mainly in response to the temperature transition occurring when the bacterium transits from the outer environment to the intestinal milieu. VirF then acts as anti-H-NS protein and directly activates the icsA and virB genes, triggering the full expression of the invasion program of Shigella. In this review we will focus on the structure of VirF, on its sophisticated regulation, and on its role as major player in the path leading from the non-invasive to the invasive phenotype of Shigella. We will address also the involvement of VirF in mechanisms aimed at withstanding adverse conditions inside the host, indicating that this protein is emerging as a global regulator whose action is not limited to virulence systems. Finally, we will discuss recent observations conferring VirF the potential of a novel antibacterial target for shigellosis

    Open-source genomic analysis of Shiga-toxin–producing E. coli O104:H4

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    An outbreak caused by Shiga-toxin–producing Escherichia coli O104:H4 occurred in Germany in May and June of 2011, with more than 3000 persons infected. Here, we report a cluster of cases associated with a single family and describe an open-source genomic analysis of an isolate from one member of the family. This analysis involved the use of rapid, bench-top DNA sequencing technology, open-source data release, and prompt crowd-sourced analyses. In less than a week, these studies revealed that the outbreak strain belonged to an enteroaggregative E. coli lineage that had acquired genes for Shiga toxin 2 and for antibiotic resistance