bIn this study, we evaluated the use of EntericBio real-time Gastro Panel I (Serosep, Limerick, Ireland) for routine use in a clinical microbiology laboratory for simultaneous detection of Campylobacter jejuni, coli, and lari, Shiga toxin-producing Escherichia coli (STEC), Salmonella spp., and Shigella spp. in feces. This system differs from its predecessor (the EntericBio Panel II system, Serosep) in that it allows real-time detection of pathogens directly from feces, without pre-enrichment. It also specifically detects Campylobacter jejuni, coli, and lari rather than all Campylobacter species, as is the case with the previous system. A total of 528 samples from patients presenting with acute gastroenteritis were screened prospectively with this assay, and results were compared with those of the current method, which combines screening the samples with a molecular assay (the EntericBio Panel II assay) and retrospective culture of the specimens in which the target was detected. Discrepancy analysis was conducted using culture and molecular methods. A cute infectious gastroenteritis is common and has important economic and social consequences for both communities and health systems due to its high rate of occurrence (1). Worldwide, it is associated with a high incidence of morbidity and mortality (2), particularly among children (3).Most clinical microbiology laboratories still use traditional, culture-based diagnostic techniques for routine detection of bacterial enteric pathogens. These are both time-consuming and laborious and, in addition, have a prolonged delay in the reporting of results. This can have a significant impact on institutions such as hospitals or nursing homes, where early detection and prevention of disease spread is crucial. Molecular detection of pathogens has, however, been shown to be faster and more sensitive than traditional culture (4, 5). It is especially important where culturebased pathogen detection is problematic or lacks sensitivity. The best example of this is the genus Campylobacter, certain members of which, such as C. jejuni, can remain viable but noncultivable (6), therefore producing false-negative results if detection is based solely on a culture. Moreover, some species which can cause gastroenteritis, such as C. upsaliensis, are unlikely to be correctly isolated and identified on common media (7). Furthermore, the exact role of some fastidious Campylobacter spp. (such as C. ureolyticus or C. concisus) in gastroenteritis is still under investigation despite their being detected in large proportions of samples from patients with gastroenteritis (8, 9).Here, we present a validation study of EntericBio real-time Gastro Panel I (Serosep, Limerick, Ireland), a commercial realtime PCR system (CE marked for in vitro use) for simultaneous detection of Campylobacter jejuni, coli, and lari, Shiga toxin-producing Escherichia coli (STEC) (through detection of Stx1 and Stx2), Shigella spp., and Salmonella spp. directly from patients' feces. MATERIALS AND METHODSPatient samples. Between ...
Campylobacter jejuni and coli are collectively regarded as the most prevalent cause of bacterial foodborne illness worldwide. An emerging species, Campylobacter ureolyticus has recently been detected in patients with gastroenteritis, however, the source of this organism has, until now, remained unclear. Herein, we describe the molecular-based detection of this pathogen in bovine faeces (1/20) and unpasteurized milk (6/47) but not in poultry (chicken wings and caeca). This is, to the best of our knowledge, the first report of the presence of this potential gastrointestinal pathogen in an animal source, possibly suggesting a route for its transmission to humans.
An investigation of the prevalence of Campylobacter ureolyticus in a variety of animals led to the identification of the strain CIT 045 T , in the faeces of captive lion-tailed macaques (Macaca silenus). Originally, believed to be Campylobacter ureolyticus based on the colony morphology and positive urease test, analysis of 16S rRNA and hsp60 gene sequences of this isolate revealed that the strain differs significantly from other species of the genus Campylobacter described to date. Species-specific primers for 16S rRNA and hsp60 genes were designed and used to identify two additional strains isolated from faeces samples from other macaques. Nucleotide sequence analysis of the 16S rRNA and hsp60 genes revealed ¡95 % and ¡82 % sequence similarity to recognized species of the genus Campylobacter respectively. All three isolates formed a distinct group within the genus Campylobacter based on their 16S rRNA and hsp60 sequences and matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) profiles. The unique species status was further supported by phenotypic characteristics of the isolates. All isolates were found to be oxidase-, catalase-and urease-positive, they grew well at 37 6C and 42 6C and produced H 2 S on TSI (triple-sugar iron) and SIM (sulfide indole motility) media. The name Campylobacter corcagiensis sp. nov. is proposed for this novel species, with the strain CIT 045 T as the type strain CIT 045 T (5LMG 27932 T , CCUG 64942 T ).The genus Campylobacter has expanded considerably since it was initially proposed by Sebald & Veron (1963), at the time of writing encompassing 24 species and eight subspecies (On, 2013). Species of the genus Campylobacter inhabit a wide variety of ecological niches and have been isolated from various sample sources and hosts; including humans, birds, livestock, pets and primates (Man, 2011). Furthermore, several of these species have been associated with diseases in humans and animals.Interestingly, even though some species of the genus Campylobacter, such as Campylobacter jejuni, have been isolated from a variety of hosts, including wild birds, domestic animals and humans, multilocus sequence typing analysis has revealed a wide genetic strain diversity of C. jejuni among different hosts (Griekspoor et al., 2013). Furthermore, some strains have been found to be hostspecific, being confined to a particular wild bird species (rather than geographical location). Other genetic populations have been found to be more widely distributed between different species and have been found to be present in humans and livestock (Griekspoor et al., 2013). This heterogeneity highlights the possibility of transspecies transmission and the zoonotic potential of many species of the genus Campylobacter.As a part of a larger study investigating possible environmental sources of Campylobacter ureolyticus, faecal samples were collected from captive wild animals. Samples were collected using sterile swabs which were immediately placed in a sterile 25 ml container filled with Bolton Broth (O...
Recently, Campylobacter ureolyticus has been detected for the first time in the faeces of patients with acute gastroenteritis using polymerase chain reaction (PCR) techniques. Cultural isolation of C. ureolyticusis is not possible using the established selective methods for the isolation of thermophilic Campylobacter spp. from faeces. The aim of the current study is to develop a new selective medium capable of isolating C. ureolyticus from faecal samples. The newly-developed medium consists of Anaerobe Basal Agar with 10 g/L additional agar, 2 g/L sodium formate and 3 g/L sodium fumarate dibasic, to which 10 mg/L nalidixic acid, 10 mg/L amphotericin B and 20 mg/L vancomycin (NAV) are added as selective agents. Validation studies have shown that this experimental selective medium completely inhibits growth of Candida spp. and of Enterococcus spp. and permits reduced growth of selected coliforms and Proteus spp. Growth of Campylobacter ureolyticus on NAV medium is optimal in anaerobic and enriched hydrogen atmospheres. Additionally, an overnight enrichment step using Bolton broth to which 2 g/L sodium formate, 3 g/L sodium fumarate dibasic and the NAV supplement are added, in place of the commercial Bolton broth supplement, allows improved recovery of C. ureolyticus from patients' faeces.
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