New 16-plex PCR method for rapid detection of diarrheagenic Escherichia coli directly from stool samples

Antikainen, Jenni; Tarkka, Eveliina; Haukka, Kaisa; Siitonen, Anja; Vaara, Martti; Kirveskari, Juha

doi:10.1007/s10096-009-0720-x

Cited by 82 publications

(72 citation statements)

References 22 publications

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“…[26][27][28][29] The increasing o pathogenic and commensal ESBL-producing E. coli strains are originated from different sources (such as human, food, and animal) and the dissemination of antibiotic resistance among other members of Enterobacteriaceae is a worldwide problem in the control and treatment of infections in human and animals. 20,[30][31][32][33][34] Moreover, some studies have shown that infections caused by ESBL-producing bacteria leads to the increase in the economic costs of infections treatment and the morbidity and mortality of diseases.…”

Section: Discussionmentioning

confidence: 99%

Phylogenetic Grouping and Phenotypic Detection of Extended-Spectrum β-Lactamases Among Escherichia coli From Calves and Dairy Cows in Khuzestan, Iran

Barzan¹,

Gharibi²,

Ghorbanpoor³

et al. 2017

Int J Enteric Pathog

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Background: Food-producing animals are under suspicion for the reservoir and colonization of ESBL (extended-spectrum beta-lactamase)-producing bacteria especially Enterobacteriaceae and therefore infection of the humans with them. The increasing reports on the ESBLs presence in the pathogenic and commensal Escherichia coli isolates have been a concern worldwide. These strains can be attributed to one of the main phylogenetic groups and subgroups. Several studies have shown the relationship between the phylogeny and antimicrobial resistance of E. coli strains. Objectives: The aim of this study was to analyze the phylogenetic group of ESBL-producing E. coli and detect its phenotype using the multiplex polymerase chain reaction (PCR) and combined disk method. Materials and Methods: Two hundred five E. coli fecal isolates were obtained from 103 calves (90 healthy and 13 diarrheic) and 102 dairy cows (healthy) from 8 farms in Khuzestan province, Iran. The triplex PCR method was used to allocate the E. coli isolates based on the presence or absence of 3 genes (chuA, yjaA, and tspE4.C2) to yield 4 definite phylogenetic groups and 7 subgroups. Phenotypic ESBL-producing E. coli was determined using the double disk diffusion method according to the manufacturer's instructions and Clinical & Laboratory Standards Institute (CLSI) guidelines. Results: A total of 65.04% and 22.3% of isolates from calves and 70.5% and 20.5% of isolates from dairy cows belonged to phylogroups B1 and A, respectively. In addition, no isolate from the diarrhoeic calves was found to belong to group B2 and subgroups D2 and A0. A low prevalence (2/205 isolates, 0.97%) of ESBL-producing E. coli was found only in the samples of dairy cows which belonged to the phylogenetic group A and phylogenetic subgroup A1. There was no statistically significant relationship between the phylogenetic group and the production of ESBLs (P = 0.11). There was also no difference between the E. coli isolates from calves and dairy cows in the production of ESBLs (P = 0.5). Conclusion: There was no statistically significant relationship between the phylogenetic group and the production of ESBLs (P = 0.11). There was also no difference between E. coli isolates from calves and dairy cows in the production of ESBLs (P = 0.5). Based on these results, there is a low prevalence of ESBL-producing E. coli in the dairy farms of Khuzestan province. However, further large scale investigations are necessary to control the antibiotic resistance in the human and animal foodstuff.

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Section: Discussionmentioning

confidence: 99%

Phylogenetic Grouping and Phenotypic Detection of Extended-Spectrum β-Lactamases Among Escherichia coli From Calves and Dairy Cows in Khuzestan, Iran

Barzan¹,

Gharibi²,

Ghorbanpoor³

et al. 2017

Int J Enteric Pathog

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“…16-plex PCR assay:The presence of STEC, EPEC, ETEC, EIEC and EAEC in human diarrhoeal stoolssamples was detected by 16-plex PCR for the genes uidA, pic, bfp, invE, hlyA, elt, ent, escV, eaeA, ipaH, aggR, stx1, stx2, estIa, estIband astA. The primers and PCR conditions were as previously described [10]. The nucleotide sequences and predicted sizes of the amplified products for the specific oligonucleotide primers used in this study are shown in Table 1.…”

Section: Cultivation Samplesmentioning

confidence: 99%

Prevalence of <i>Escherichia coli</i> virulence genes in patients with diarrhoea in Ouagadougou, Burkina Faso

Somda¹,

Bonkoungou²,

Zongo³

et al. 2017

Af J Clin Exp Micro

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Objective: Diarrhoeagenic E. coli (DEC) strains are important causes of diarrhoea in the developing world and, to a lesser extent, inthe developed world. In this study, we investigated the prevalence of the virulence genes specific for five major pathogroups of diarrheagenic Escherichia coli (DEC) in primary cultures from diarrhoeagenic patients in Burkina Faso.Methodology: From September 2016 to Mars 2017, a total of 211 faecal samples from diarrhoeagenic patients from urban hospitals of Ouagadou, Burkina Faso have been analysed. A 16-plex PCR was used to detect simultaneously, the five major DEC pathotypes (enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), Shiga toxin-producing E. coli (STEC), enteroaggregative E. coli (EAEC) and enteroinvasive E. coli (EIEC)). Results: At least one diarrhoeagenic E. Coli pathotype was detected in 31 samples (14.7%) in children and adults with diarrhoea. EAEC was the most common pathotype detected 9.5% (20/211), followed by EIEC2.4% (05/211) and STEC 0.5% (01/211). More than one DEC pathotype were detected in 2.4% (05/211) patients. EPEC and ETEC were not detected in single infection but in co-infection with others pathotypes. Conclusion: DEC, especially enteroaggregative, may be important responsible of diarrhoeas in Burkina Faso from all ages patient.Key Words: Diarrhoeagenic Escherichia coli, 16-plex PCR, Burkina Faso, human diarrhoeas stool. PREVALENCE DES GENES DE VIRULENCE D'ESCHERICHIA COLIISOLEES DES SELLES DIARRHEIQUES CHEZ LES PATIENTS

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“…Therefore, rapid and reliable detection of STEC isolates in food and clinical samples is required. The direct molecular detection of Shiga toxin (Stx1 or Stx2) genes alone or in combination with other genes coding for major STEC virulence factors by nucleic acid-based techniques is known to be highly sensitive and specific in STEC detection (1,2,10,25). Major disadvantages of these assays are, however, the labor-intensive sample preparation and the increase of the costs when used only for screening purposes.…”

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confidence: 99%

Usability and Performance of CHROMagar STEC Medium in Detection of Shiga Toxin-Producing Escherichia coli Strains

2012

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The performance and usability of CHROMagar STEC medium (CHROMagar Microbiology, Paris, France) for routine detection of Shiga toxin-producing Escherichia coli (STEC) strains were examined. The ability of the medium to selectively propagate STEC strains differing by their serotypes and virulence genes was studied with a collection of diarrheagenic E. coli isolates (n ‫؍‬ 365) consisting of 49 different serotypes and with non-STEC and other bacterial isolates (n ‫؍‬ 264). A total of 272 diarrheagenic E. coli (75.0%) isolates covering 24 different serotypes grew on CHROMagar STEC. The highest detection sensitivities were observed within the STEC serogroups O26 (90.0%), O111 (100.0%), O121 (100.0%), O145 (100.0%), and O157 (84.9%), and growth on CHROMagar STEC was highly associated with the presence of the tellurite resistance gene (terD). The specificity of the medium was 98.9%. In addition, CHROMagar STEC was used in parallel with a Shiga toxin-detecting immunoassay (Ridaquick Verotoxin/O157 Combi; R-biopharm, Darmstadt, Germany) to screen fecal specimens (n ‫؍‬ 47) collected from patients suffering from hemorrhagic diarrhea. Positive growth on CHROMagar STEC was confirmed by the Premier EHEC enzyme immunoassay (Meridian Bioscience, Inc., Cincinnati, OH), and discrepant results between the two screening methods were confirmed by stx gene-detecting PCR. All 16 of the 47 stool samples that showed positive growth on CHROMagar STEC were also positive in the confirmatory tests. CHROMagar STEC proved to be an interesting option for STEC screening, allowing good detection sensitivity and specificity and permitting strain isolation for further outbreak investigations when required. Shiga toxin-producing Escherichia coli (STEC) strains are frequently identified as causative agents of a wide spectrum of diseases, ranging from mild gastroenteritis and hemorrhagic colitis to life-threatening diseases such as hemolytic-uremic syndrome (HUS) (14). Due to their pathogenic properties, particularly phage-encoded Shiga toxins (Stxs), these strains are also called enterohemorrhagic E. coli (EHEC) (22).Although sorbitol-negative serotype O157:H7 has been the most common STEC serotype, the clinical significance of non-O157 STEC strains is on the rise worldwide (11,12,14,15). STEC strains are known to be diverse, and in addition to classical EHEC strains, newly emerging strains have recently been associated with severe clinical illness and with outbreaks in Europe (7,17,18). Moreover, there are reports that non-O157 STEC strains have been found in foods retailed for consumption in raw form by humans (4,5,19).The increasing prevalence of these intestinal pathogens has important public health implications. Therefore, rapid and reliable detection of STEC isolates in food and clinical samples is required. The direct molecular detection of Shiga toxin (Stx1 or Stx2) genes alone or in combination with other genes coding for major STEC virulence factors by nucleic acid-based techniques is known to be highly sensitive and specific in STEC dete...

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New 16-plex PCR method for rapid detection of diarrheagenic Escherichia coli directly from stool samples

Cited by 82 publications

References 22 publications

Phylogenetic Grouping and Phenotypic Detection of Extended-Spectrum β-Lactamases Among Escherichia coli From Calves and Dairy Cows in Khuzestan, Iran

Phylogenetic Grouping and Phenotypic Detection of Extended-Spectrum β-Lactamases Among Escherichia coli From Calves and Dairy Cows in Khuzestan, Iran

Prevalence of <i>Escherichia coli</i> virulence genes in patients with diarrhoea in Ouagadougou, Burkina Faso

Usability and Performance of CHROMagar STEC Medium in Detection of Shiga Toxin-Producing Escherichia coli Strains

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