Evaluation of a PCR detection method for Escherichia coli O157:H7/H- bovine faecal samples

Four commercial DNA extraction methods, PrepMan Ultra (Applied Biosystems), InstaGene Matrix (BioRad), DNeasy Tissue kit (Qiagen), and UltraClean (MoBio), were tested for PCR detection of Listeria monocytogenes, Escherichia coli O157: H7, Salmonella, and Staphylococcus aureus in fresh, minimally processed vegetables. For comparative purposes, sensitivity assays with specific PCRs were carried out after DNA extraction with the four methods in green pepper, broccoli, and onion artificially inoculated with the four pathogens separately. As confirmed by statistical analysis, the DNeasy Tissue kit rendered the highest sensitivity values in the three matrices assayed for Salmonella, L. monocytogenes, and E. coli O157:H7 and in onion for S. aureus. Despite being the most expensive of the methods compared, the DNeasy Tissue Kit can be successfully applied for any of the four most commonly studied pathogens, thus saving time and overall reducing the cost of the analysis.

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Sorbitol-fermenting Shiga toxin-producing Escherichia coli O157: indications for an animal reservoir

Epidemiology and Infection ◽

10.1017/s0950268805005467 ◽

2005 ◽

Vol 134 (4) ◽

pp. 719-723 ◽

Cited By ~ 25

Author(s):

D. ORTH ◽

K. GRIF ◽

M. P. DIERICH ◽

R. WÜRZNER

Keyword(s):

Escherichia Coli ◽

Clinical Diagnosis ◽

Shiga Toxin ◽

Haemolytic Uraemic Syndrome ◽

Stool Specimen ◽

Farm Animals ◽

Molecular Characteristics ◽

Escherichia Coli O157 ◽

Animal Reservoir ◽

Faecal Samples

This study investigates a sorbitol-fermenting enterohaemorrhagic Escherichia coli (SF EHEC) O157 infection in a farmer's family in the Austrian province of Salzburg. The investigation commenced after a 10-month-old boy was admitted to hospital with the clinical diagnosis of a haemolytic–uraemic syndrome (HUS) and his stool specimen grew SF EHEC O157:H−. In a subsequent environmental survey, a stool specimen of the 2-year-old brother and faecal samples of two cattle from the family's farm were also found to be positive for SF EHEC O157:H−. All four isolates had indistinguishable phenotypic and molecular characteristics and were identical to the first strain detected in Bavaria in 1988. Despite identical isolates being demonstrated in Bavaria after 1988, and until this report, increased surveillance in neighbouring Austria had not found this organism. We propose that the strain may have recently spread from Bavaria to Austria. Although SF EHEC O157:H− strains are still rare, they may represent a considerable health threat as they can spread from farm animals to humans and between humans.

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First isolation of Escherichia coli O157:H7 from faecal and milk specimens from Anatolian water buffaloes (Bubalus bubalus) in Turkey

Journal of the South African Veterinary Association ◽

10.4102/jsava.v79i4.267 ◽

2008 ◽

Vol 79 (4) ◽

Cited By ~ 6

Author(s):

E. Seker ◽

H. Yardimci

Keyword(s):

Escherichia Coli ◽

Raw Milk ◽

Escherichia Coli O157 ◽

Water Buffaloes ◽

E Coli ◽

Milk Samples ◽

Cultural Methods ◽

Faecal Samples

Three hundred rectal faecal samples and 213 raw milk samples obtained from the tanks and containers were examined using standard cultural methods. Escherichia coli O157:H7 was isolated from 11 (3.7 %) of 300 faecal samples and 3 (1.4 %) of 213 raw milk samples. It was determined that 8 (73 %) of E. coli O157:H7 strains isolated from faecal samples originated from water buffaloes younger than 2 years of age and 3 (27 %) from 2-year-old and older water buffaloes. This is the 1st isolation of Escherichia coli O157:H7 from faecal and milk samples of water buffaloes in Turkey.

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A Study on Prevalence of Escherichia coli O157 with a Verified Method in Foods

Turkish Journal of Agriculture - Food Science and Technology ◽

10.24925/turjaf.v5i4.294-298.1005 ◽

2017 ◽

Vol 5 (4) ◽

pp. 294 ◽

Cited By ~ 1

Author(s):

Semiha Yalçın ◽

Ayla Ünver Alçay ◽

Gözde Yüzbaşıoğlu ◽

Burcu Çakmak ◽

Aysun Sağlam

Keyword(s):

Escherichia Coli ◽

Detection Limit ◽

Standard Method ◽

False Positive ◽

Detection Method ◽

Minimum Detection Limit ◽

Escherichia Coli O157 ◽

E Coli

The purpose of this study were to identify the presence of E.coli O157 and to determine its prevalence in foods which were collected from various restaurants, shops and markets in Istanbul. Also, validation of detection method of E. coli O157 in all food stuffs was carried out according to applicability, repeatability, and minimum detection limit (LOD) and false positive and negative analysis based on TS EN ISO 16654 standard method. The results showed that the prevalence of E. coli O157 in food was 2%, and its prevalence increased in April and May.

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Development of a rapid capture-cum-detection method for Escherichia coli O157 from apple juice comprising nano-immunomagnetic separation in tandem with surface enhanced Raman scattering

International Journal of Food Microbiology ◽

10.1016/j.ijfoodmicro.2014.07.036 ◽

2014 ◽

Vol 189 ◽

pp. 89-97 ◽

Cited By ~ 32

Author(s):

Roya Najafi ◽

Shubhasish Mukherjee ◽

Jim Hudson ◽

Anup Sharma ◽

Pratik Banerjee

Keyword(s):

Escherichia Coli ◽

Raman Scattering ◽

Apple Juice ◽

Detection Method ◽

Surface Enhanced Raman Scattering ◽

Immunomagnetic Separation ◽

Escherichia Coli O157 ◽

Surface Enhanced ◽

Surface Enhanced Raman ◽

Enhanced Raman Scattering

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The occurrence and sources of Campylobacter spp., Salmonellaenterica and Escherichia coli O157:H7 in the Salmon River, British Columbia, Canada

Journal of Water and Health ◽

10.2166/wh.2009.076 ◽

2009 ◽

Vol 8 (2) ◽

pp. 374-386 ◽

Cited By ~ 42

Author(s):

Cassandra C. Jokinen ◽

Hans Schreier ◽

William Mauro ◽

Eduardo Taboada ◽

Judith L. Isaac-Renton ◽

...

Keyword(s):

Escherichia Coli ◽

British Columbia ◽

Water Samples ◽

Escherichia Coli O157 ◽

Salmon River ◽

E Coli ◽

Faecal Samples ◽

Precipitation Analysis ◽

Species Markers ◽

Pcr Techniques

In this study, we wished to assess the prevalence and determine the sources of three zoonotic bacterial pathogens (Salmonella, Campylobacter, and Escherichia coli O157:H7) in the Salmon River watershed in southwestern British Columbia. Surface water, sewage, and animal faecal samples were collected from the watershed. Selective bacterial culture and PCR techniques were used to isolate these three pathogens and indicator bacteria from these samples and characterize them. Campylobacter was the most prevalent pathogen in all samples, followed by Salmonella, and E. coli O157:H7. E. coli O157:H7 and Salmonella isolation rates from water, as well as faecal coliform densities correlated positively with precipitation, while Campylobacter isolation rates correlated negatively with precipitation. Analysis of DNA extracted from water samples for the presence of Bacteroides host-species markers, and comparisons of C. jejuniflaA-RFLP types and Salmonella serovars from faecal and water samples provided evidence that human sewage and specific domestic and wild animal species were sources of these pathogens; however, in most cases the source could not be determined or more than one source was possible. The frequent isolation of these zoonotic pathogens in the Salmon River highlights the risks to human health associated with intentional and unintentional consumption of untreated surface waters.

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