Search results for "S.aureus"

PCR-based procedures for detection and quantification of Staphylococcus aureus and their application in food.

2006

Aims:  To evaluate the specificity of nuc targeted primers for PCR detection of Staphylococcus aureus in different food matrices and to establish a RTQ-PCR procedure suitable for the routine detection and quantification of this pathogen in food. Methods and Results:  Specificity of nuc targeted primers (Pri1–Pri2 and the newly designed RTQ-PCR primers) was tested on a total of 157 strains of genetically confirmed identity, including reference and food isolates. PCR detection on artificially inoculated beef samples by DNA extraction using a DNeasy Tissue Kit (Qiagen GmhH, Hilden, Germany) showed a sensitivity value around 103 CFU g−1. The two RTQ-PCR systems, incorporating SYBR-Green I and T…

Identification and typing of food-borne Staphylococcus aureus by PCR-based techniques.

2005

Abstract The possibility of using PCR for rapid identification of food-borne Staphylococcus aureus isolates was evaluated as an alternative to the API-Staph system. A total of 158 strains, 15 S. aureus , 12 other staphylococcal species, and 131 isolates recovered from 164 food samples were studied. They were phenotypically characterized by API-Staph profiles and tested for PCR amplification with specific primers directed to thermonuclease ( nuc ) and enterotoxin ( sea to see ) genes. Disagreement between the PCR results and API-Staph identification was further assessed by the analysis of randomly amplified polymorphic DNA (RAPD) profiles obtained with three universal primers (M13, T3, and T…

Real-time quantitative PCR of Staphylococcus aureus and application in restaurant meals.

2006

Staphylococcus aureus is considered the second most common pathogen to cause outbreaks of food poisoning, exceeded only by Campylobacter. Consumption of foods containing this microorganism is often identified as the cause of illness. In this study, a rapid, reliable, and sensitive real-time quantitative PCR was developed and compared with conventional culture methods. Real-time quantitative PCR was carried out by purifying DNA extracts of S. aureus with a Staphylococcus sample preparation kit and quantifying it in the LightCycler system with hybridization probes. The assay was linear from a range of 10 to 10(6) S. aureus cells (r2 > 0.997). The PCR reaction presented an efficiency of >85%. …

Concomitant loss of conformation and superantigenic activity of staphylococcal enterotoxin B deletion mutant proteins.

1993

The T-cell-stimulating activity of staphylococcal enterotoxin B (SEB) is an important factor in the pathogenesis of certain staphylococcal diseases. To investigate the immunologically active domains of the SEB molecule, we have produced truncated fragments of recombinant SEB by C-terminal and N-terminal deletions. The fragments were expressed as fusion proteins with protein A, including a cleavage site to remove the protein A part. Mutant proteins were tested for the ability to stimulate human resting T cells and SEB-reactive T-cell clones. Deletion of only 9 amino acids from the C terminus leads to complete loss of T-cell-stimulating activity. Removing further amino acids from the SEB mole…

Comparison of Four Commercial DNA Extraction Kits for PCR Detection of Listeria monocytogenes, Salmonella, Escherichia coli O157:H7, and Staphylococc…

2008

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. monocytoge…

Rapid whole protein quantification of staphylococcal enterotoxin B by liquid chromatography

2012

Abstract Food poisoning caused by Staphylococcus aureus is one of the most important foodborne diseases in the world. The ability of these bacteria to produce one or more enterotoxins in milk and dairy products is linked to staphylococcal food poisoning. Enterotoxin B (SEB) is an exotoxin produced by S. aureus and is one of the compounds most frequently involved in staphylococcal food poisoning worldwide. In this work, 20 samples of milk collected from restaurants have been studied for the presence of S. aureus enterotoxigenic strains. All the isolates from milk samples have been analysed by liquid chromatography-coupled with diode array detector for the rapid identification and quantificat…

Antibacterial Activity of Extracts from Some Bryophytes

2012

The antimicrobial activity of aqueous and ethanolic extracts of 11 Bryophyta species and 9 Marchantiophyta species collected in Latvia was tested against Staphylococcus aureus, Escherichia coli and Bacillus cereus. The extract of Lophocolea heterophylla inhibited the growth of B. cereus, but none of the tested extracts inhibited the growth of E. coli. 70% of bryophyte species demonstrated certain activity in relation to S. aureus. In general, 73% of ethanolic extracts and 39% of aqueous extracts exhibited antibacterial activity against S. aureus. The highest degree of antibacterial activity against S. aureus was shown by the ethanolic extract of Dicranum scoparium and aqueous extracts of At…

Staphyloccal alpha toxin

1998

Viability of microorganisms in novel antineoplastic and antiviral drug solutions

1998

Introduction. In determining the expiration-dates of ready-to-use antineoplastic and antiviral drug solu tions, microbiological aspects must be considered. This is especially true because many antineoplastic drugs introduced into the market are already known to lack antimicrobial activity. The purpose of this study is to evaluate the growth of four different microorganisms in ready-to-use solutions of 14 differ ent novel antineoplastic and antiviral drugs. Methods. The lowest concentrations of 14 dif ferent antineoplastic and antiviral drugs prescribed in our hospital were prepared in polyvinyl chloride bags or a polyethylene container (paclitaxel) containing 0.9% sodium chloride or 5% dex…

Coincidental loss of bacterial virulence in multi-enemy microbial communities.

2014

The coincidental virulence evolution hypothesis suggests that outside-host selection, such as predation, parasitism and resource competition can indirectly affect the virulence of environmentally-growing bacterial pathogens. While there are some examples of coincidental environmental selection for virulence, it is also possible that the resource acquisition and enemy defence is selecting against it. To test these ideas we conducted an evolutionary experiment by exposing the opportunistic pathogen bacterium Serratia marcescens to the particle-feeding ciliate Tetrahymena thermophila, the surfacefeeding amoeba Acanthamoeba castellanii, and the lytic bacteriophage Semad11, in all possible combi…