BACKGROUND: We investigated the prevalence of various pathogens (13 enteric bacteria and 5 viruses) which cause diarrhea using multiplex PCR of stool specimens and compared two multiplex PCR methods for detecting diarrheagenic Escherichia coli. METHODS: A total of 405 stool specimens submitted between November 2010 to February 2011 for routine culture of enteric pathogens were included and screened for five viruses (astrovirus, Group A rotavirus, enteric adenovirus, norovirus G1/G2) and eight bacteria (Salmonella spp., Shigella spp., Campylobacter spp., Vibrio spp., C. difficile Toxin B, C. perfringens, Y. enterolytica, Aeromonas spp.) using the Seeplex(R) Diarrhea ACE detection kit (Seegene). In addition, virulence-associated genes of enteropathogenic E. coli, (EPEC), enterohemorrhagic E. coli (EHEC), enteroinvasive E. coli, (EIEC), enterotoxigenic E. coli (ETEC), and enteroaggressive E. coli (EAEC) were detected using 16-plex PCR and a commercial diarrheagenic E. coli detection (DEC) PCR kit (SSI Diagnostica). RESULTS: Overall, 138 (34.1%) of 405 samples was positive for pathogen. The positive rate for virus was 18.5%. norovirus G2, Group A rotavirus, enteric adenovirus, astrovirus and norovirus G1 were detected in 40, 23, 8, 3 and 1 samples, respectively. The positive rate for bacteria was 24.4% (99/405). C. difficile toxin B was the most frequently detected, followed by C. perfringens, EPEC, and EAEC. The agreements of the two multiplex PCR methods for detecting EPEC and EHEC were 99.3% and 100%, respectively. CONCLUSION: The detection rate was high (34.1%) including various diarrheagenic E. coli (6.2%) and C. perfringens (5.2%). Multiplex PCR is thus useful for detecting various pathogens.
Adenoviridae ; Aeromonas ; Bacteria ; Campylobacter ; Diarrhea ; Enterobacteriaceae ; Enterohemorrhagic Escherichia coli ; Enteropathogenic Escherichia coli ; Enterotoxigenic Escherichia coli ; Escherichia ; Escherichia coli ; Multiplex Polymerase Chain Reaction ; Norovirus ; Polymerase Chain Reaction ; Prevalence ; Rotavirus ; Shigella ; Vibrio ; Viruses

Verotoxin-producing Escherichia coli O157 is a primary cause of severe and bloody diarrhea. Campylobacter spp. are one of the commonly reported bacterial cause of gastrointestinal infections throughout the world. Only a few cases involving both E. coli O157 and Campylobacter species have been reported. The authors simultaneously isolated verotoxin-producing E. coli O157 and Campylobacter species from the stool of a 3 year-old male with bloody diarrhea, fever and abdominal pain.
Abdominal Pain ; Campylobacter* ; Child, Preschool ; Diarrhea ; Escherichia coli O157* ; Escherichia coli* ; Escherichia* ; Fever ; Gastroenteritis* ; Humans ; Male ; Shiga Toxins ; Shiga-Toxigenic Escherichia coli

BACKGROUND: The Polymerase Chain Reaction (PCR) for the Shiga toxin has been widely used for diagnosis of Shiga toxin-producing Escherichia coli (STEC) infection including Escherichia coli O157 (O157) instead of using a culture. However, bacteriological isolation must be followed for final diagnosis. Our study was aimed to compare the detection limit between the culture after the acid treatment and the PCR after enrichment. METHODS: The standard strain of O157 was cultured, diluted and mixed with the stool of normal adult in order to make a final concentration of the 10(1)-10(5) colony forming unit (CFU)/g of stool. Each concentration of samples was enriched in a trypticase soy broth for 6 hours at 42degrees C and treated with acid to suppress normal flora. Then it was streaked on cefixime-tellurite-sorbitol MacConkey (CT-SMAC) agar evenly and cultured for 18 hours at 37degrees C. The same concentrations of bacterial suspension in the stool were enriched in a Luria-Bertani (LB) broth overnight at 37degrees C. The centrifuged pellets from 1 mL of each concentration of the samples were boiled and DNA was extracted using the resin method and PCR was performed to amplify stx2. RESULTS: The detection limit for the culture after acid treatment was 10(3) CFU/g of the stool and that for PCR after enrichment was 101 CFU/g of the stool. CONCLUSIONS: Culture after acid treatment for O157 would be an effective method for isolation of O157 from a patient's stool. However, this method is less sensitive than the PCR after enrichment as far as detection limit is concerned. A combination of both methods would be an effective method for detecting O157 from patient stools.
Adult ; Agar ; Diagnosis ; DNA ; Escherichia coli O157* ; Escherichia coli* ; Escherichia* ; Humans ; Limit of Detection ; Polymerase Chain Reaction* ; Shiga Toxin ; Shiga-Toxigenic Escherichia coli ; Stem Cells

Escherichia coli O157 is an important serotype of enterohemorrhagic E. coli that causes hemorrhagic colitis worldwide. Outbreaks of E. coli O157 have been assocoated with contaminated food like meat, raw milk, and water, but recently vegetables and fruits have accounted for a growing number of recognized outbreaks. We isolated verotoxin producing E. coli O157 from the stool of a 3 year-old female with bloody diarrhea and abdominal pain. The child had been eating salad with vegetables and fruits frequently.
Abdominal Pain ; Child ; Colitis ; Diarrhea ; Disease Outbreaks ; Eating ; Enterohemorrhagic Escherichia coli ; Escherichia ; Escherichia coli ; Escherichia coli O157 ; Female ; Fruit ; Humans ; Meat ; Milk ; Shiga Toxins ; Vegetables

Attaching and effacing Escherichia coli (AEEC) cause enteric infections in humans and animals. Attaching indicates the intimate attachment of bacteria to the enterocyte, and effacing relates to the localized effacement of brush border microvilli. Enteropathogenic (EPEC) and enterohemorrhagic Escherichia coli (EHEC) infections are characterized by the formation of attaching and effacing (AE) lesion on the intestinal epithelial cells. Therefore, they are often grouped together as AEEC. Development of multiplex PCR allowed us to type five of the most important genes implicated in the formation of the AE lesion. A total of 60 AEEC strains isolated from diarrheal patients were investigated by multiplex PCR for the presence of the insertion site of locus of enterocyte effacement (LEE) and LEE-related (eae, tir, espA, espB, and espD) genes. Associating the results of LEE genes typing in the AEEC strains, three different pathotypes are determined: eae(gamma)-tir(gamma)-espA(gamma)-espB(gamma)-espD(gamma) (O157:H7), eae(beta)-tir(beta)-espA(beta)-espB(beta)-espD(beta) (O26:H11), and eae(alpha)-tir(alpha)-espA(alpha)-espB(alpha)-espD(alpha) (O55:H6). These results indicate that AEEC are a heterogenous groups of organisms.
Animals ; Bacteria ; Enterocytes* ; Enterohemorrhagic Escherichia coli* ; Enteropathogenic Escherichia coli* ; Epithelial Cells ; Escherichia coli ; Humans ; Microvilli ; Multiplex Polymerase Chain Reaction

Enterohemorrhagic Escherichia coli serotype O157:H7 is a pathotype of diarrheagenic E. coli that produces one or more Shiga toxins, forms a characteristic histopathology described as attaching and effacing lesions, and possesses the large virulence plasmid pO157. The bacterium is recognized worldwide, especially in developed countries, as an emerging food-borne bacterial pathogen, which causes disease in humans and in some animals. Healthy cattle are the principal and natural reservoir of E. coli O157:H7, and most disease outbreaks are, therefore, due to consumption of fecally contaminated bovine foods or dairy products. In this review, we provide a general overview of E. coli O157:H7 infection, especially focusing on the bacterial characteristics rather than on the host responses during infection.
Animals ; Cattle ; Cattle Diseases/blood ; Cattle Diseases/epidemiology ; Developing Countries ; Enterohemorrhagic Escherichia coli ; Escherichia coli Infections/blood ; Escherichia coli Infections/epidemiology ; Escherichia coli Infections/veterinary ; Escherichia coli O157/genetics ; Escherichia coli O157/pathogenicity ; Feces/microbiology ; Hemolytic-Uremic Syndrome/blood ; Hemolytic-Uremic Syndrome/epidemiology ; Hemolytic-Uremic Syndrome/veterinary ; Operon ; Shiga Toxins/analysis ; Shigella dysenteriae ; Virulence

No abstract available.
Enterotoxigenic Escherichia coli ; Enterotoxins

No abstract available.
Enterotoxigenic Escherichia coli

No abstract available.
Enterotoxigenic Escherichia coli ; Enterotoxins

No abstract available.
Enterotoxigenic Escherichia coli