Detection of Escherichia coli O157:H7 by multiplex PCR

Fratamico, Pina M.; Sackitey, Solomon; Deng, Ming

doi:10.1128/jcm.33.8.2188-2191.1995

Cited by 187 publications

(73 citation statements)

References 25 publications

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“…gasseri K7, (Shu et al 1999;Galdeano and Perdigon 2004;Matijasic et al 2004Matijasic et al , 2006bPaturi et al 2007) (Torriani et al 1999). Colonies grown on E coli O157:H7 agar were identified by multiplex PCR (Fratamico et al 1995).…”

Section: Feeding Infection Experimentsmentioning

confidence: 99%

Lactobacillus gasseriK7 modulates the blood cell transcriptome of conventional mice infected withEscherichia coliO157:H7

et al. 2014

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Aims: As the immune cells underlying the intestinal barrier sense luminal microbial signals, blood cell transcriptomics may identify subclinical changes triggered by gut bacteria that may otherwise not be detected. We have therefore investigated how Lactobacillus gasseri K7 and enterohemorrhagic Escherichia coli O157:H7 modulate the blood cell transcriptome of mice possessing an intact microbiota. Methods and Results:We have analysed the transcriptome of five groups of C57BL/6J mice: (i) control, (ii) inoculated with a single dose of E. coli, (iii) inoculated during 2 weeks with Lact. gasseri, (iv) co-inoculated with E. coli and Lact. gasseri, (v) inoculated with Lact. gasseri prior to E. coli infection. The transcriptome could distinguish between the five treatment groups. Gene characteristics of bacterial infection, in particular inflammation, were upregulated in the mice inoculated with E. coli. Lact. gasseri had only mild effects on the transcriptome but modified the gene expression induced by E. coli. Conclusions: The transcriptome differentiates mice inoculated orally with E. coli, Lact. gasseri and combinations of these two strains. Significance and Impact of the Study: These results suggest that the blood cell transcriptome can be used as a source of biomarkers to monitor the impact of probiotics in subclinical models of infectious disease.

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Section: Feeding Infection Experimentsmentioning

confidence: 99%

Lactobacillus gasseriK7 modulates the blood cell transcriptome of conventional mice infected withEscherichia coliO157:H7

et al. 2014

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“…The day after death, the three dead animals were submitted to the laboratory for necroscopy, routine bacteriology, virology, parasitology and histology. Briefly, two intestinal swabs (jejunum and colon) and one swab from spleen, liver and brain were collected and cultured on MacConkey agar, 5% blood sheep agar and Mannitol Salt Agar, incubated at 37 8C for 24 h; Escherichia coli isolates were further tested for eae, stx1, stx2, sta, stb, lt and hle genes by PCR (Gannon et al, 1992;Fratamico et al, 1995;Gannon et al, 1997;Osek, 1999). The presence of rotavirus and coronavirus in intestinal content was assessed by transmission electron microscopy.…”

Section: Case Reportmentioning

confidence: 99%

Severe weight loss in lambs infected with Giardia duodenalis assemblage B

Aloisio

Filippini

Antenucci³

et al. 2006

Veterinary Parasitology

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An outbreak of giardiasis was observed in a sheep farm in Central Italy. Infected lambs (30-90 days of age) showed a malabsorption syndrome, decreased weight gain and impairment in feed efficiency. The most relevant clinical sign was the excretion of malodorous and poorly formed faeces, whereas diarrhoea was rarely observed in the flock. Laboratory investigations revealed the presence of Giardia in affected animals, while no other significant viral, bacterial or parasitic pathogens were identified in faeces or tissue samples. A mild to severe infiltrative enteritis with eosinophils, lymphocytes and plasma cells was detected in histological sections of the gut. Giardia parasites collected from duodenal aspirates were typed as Giardia duodenalis Assemblage B, by PCR amplification and sequencing of the TPI gene. Treatment with fenbendazole at a dose of 10mg/kg for 3 consecutive days, successfully cleared the infection. These results show that G. duodenalis can cause significant economic losses in sheep farming.

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“…In particular, the polymerase chain reaction (PCR) has been used successfully for the detection of Escherichia coli O157:H7, Listeria monocytogenes and Bacillus spp. in milk and other dairy products (Lantz et al 1994a;Candrian 1995;Dickinson et al 1995;Fratamico et al 1995;Herman et al 1995;Tsen et al 1996;Herman et al 1997). Many food components, however, are inhibitory to PCR, by either interfering with cell lysis (i.e., DNA extraction), contributing to nucleic acid degradation or inhibiting Taq DNA polymerase (Rossen et al 1992;Herman and De Ridder 1993;Powell et al 1994;Bickley et al 1996;Wilson 1997).…”

Section: Introductionmentioning

confidence: 99%

A comparison of methods for the detection of Escherichia coli O157:H7 from artificially-contaminated dairy products using PCR

2000

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2000. Rapid nucleic acid-based methods to detect human pathogens in foods are dependent on the reliability of the DNA or RNA extraction method used. Skim milk, non-fat dry milk, Cheddar and Brie cheese, and reconstituted whey powder were seeded with serially diluted (10 0 À10 7 cfu 10 ml À1 ) Escherichia coli O157:H7 and subjected to DNA extraction (i) directly from the food product using a solvent-based procedure and (ii) using a guanidinium isothiocyanate (GITC) procedure after previous bacterial concentration. Both the ef®ciency of DNA extraction and the overall PCR detection limits were evaluated. In almost all instances, the total DNA yield using the solvent method was greater than that obtained for the concentration method. However, the purity of the DNA obtained after bacterial concentration was signi®cantly better than that obtained after organic extraction alone. PCR detection limits after each DNA recovery method varied with the speci®c food, ranging from 10 1 to 10 4 cfu ml À1 for all products except whey powder. DNA yields and subsequent PCR detection limits for reconstituted whey powder were extremely poor, and neither procedural changes nor the addition of PCR enhancement agents were able to improve recovery and/or detection. It is concluded that the ef®ciency of DNA extraction is an extremely important and frequently overlooked variable impacting the overall detection limits of PCR-based detection strategies.

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Detection of Escherichia coli O157:H7 by multiplex PCR

Cited by 187 publications

References 25 publications

Lactobacillus gasseriK7 modulates the blood cell transcriptome of conventional mice infected withEscherichia coliO157:H7

Lactobacillus gasseriK7 modulates the blood cell transcriptome of conventional mice infected withEscherichia coliO157:H7

Severe weight loss in lambs infected with Giardia duodenalis assemblage B

A comparison of methods for the detection of Escherichia coli O157:H7 from artificially-contaminated dairy products using PCR

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