Search results for "Food Contamination"
showing 10 items of 280 documents
Nanomaterials and new biorecognition molecules based surface plasmon resonance biosensors for mycotoxin detection
2019
Mycotoxins are highly toxic secondary metabolites, which may contaminate many types of food and feeds. These toxins have serious health risks for both human and animals. One of the effective ways to prevent food contamination and protect people against mycotoxins is based on timely detection. Several methods like enzyme-linked immunosorbent assay and affinity chromatography are commercially available for this purpose. Nevertheless, sensitive, fast, simple, low-cost, and portable devices are absolutely required for a fast point-of care information and making decisions. Application of biosensors appears to be a possible technique to meet this need for mycotoxins analyze. The present study has…
Control of pesticide residues by liquid chromatography-mass spectrometry to ensure food safety.
2006
Liquid chromatography-mass spectrometry (LC-MS) has become an invaluable technique for the control of pesticide residues to ensure food safety. After an introduction about the regulations that highlights its importance to meet the official requirements on analytical performance, the different mass spectrometers used in this field of research, as well as the LC-MS interfaces and the difficulties associated with quantitative LC-MS determination, are discussed. The ability to use practical data for quantifying pesticides together with the option of obtaining structural information to identify target and non-target parent compounds and metabolites are discussed. Special attention is paid to the…
Quantification of Listeria monocytogenes in salads by real time quantitative PCR
2005
Abstract A real time quantitative PCR (RTQ-PCR) was carried out purifying DNA extracts of Listeria monocytogenes using a High Pure Listeria Sample Preparation Kit and quantifying in a LightCycler system with hybridisation probes. A standard curve was constructed with serial dilutions. A range linear relationship, from 10 to 10 5 L. monocytogenes colony forming units (CFU), was observed between threshold cycle ( C t ) and logarithmic concentration of the serial dilutions. The assay was linear in a range from 10 to 10 5 L. monocytogenes CFU and the coefficient of determination ( r 2 ) was > 0.98. RTQ-PCR presented an efficiency of > 85%. The accuracy of the PCR-based assay, expressed as % bia…
A multiplex RTi-PCR reaction for simultaneous detection of Escherichia coli O157:H7, Salmonella spp. and Staphylococcus aureus on fresh, minimally pr…
2007
In this work, a new multiplex single-tube real-time PCR approach is presented for the detection of Escherichia coli O157:H7, Salmonella spp. and Staphylococcus aureus, three of the more frequent food-borne bacterial pathogens that are usually investigated in a variety of food matrices. The study includes the design and specificity testing, of a new primer and probe specific for Salmonella spp. Reaction conditions were adjusted for the simultaneous amplification and detection of specific fragments in the beta-glucuronidase (uidA, E. coli) and Thermonulease (nuc, Sta. aureus) genes, and in the replication origin sequence (oriC, Salmonella spp.). Melting-curve analysis using a SYBR Green I RTi…
Use of a species-specific multiplex PCR for the identification of pediococci.
2008
In this study, the 23S rRNA genes of nine different Pediococcus type strains were sequenced. By using a multiple sequence alignment with 23S rDNA sequences of related lactic acid bacteria two primer pairs were constructed, one for the general identification of the genus Pediococcus and one for the identification of the atypical species, P. dextrinicus. Furthermore, a primer set for a rapid multiplex PCR identification of the eight typical Pediococcus species was developed. With this technique, the species P. damnosus, P. parvulus, P. inopinatus, P. cellicola, P. pentosaceus, P. acidilactici, P. claussenii, and P. stilesii could be discriminated simultaneously in a single PCR. Experiments wi…
pilF polymorphism-based real-time PCR to distinguish Vibrio vulnificus strains of human health relevance
2012
The Gram-negative bacterium Vibrio vulnificus is a common inhabitant of estuarine environments. Globally, V. vulnificus is a significant foodborne pathogen capable of causing necrotizing wound infections and primary septicemia, and is a leading cause of seafood-related mortality. Unfortunately, molecular methods for the detection and enumeration of pathogenic V. vulnificus are hampered by the genetically diverse nature of this pathogen, the range of different biotypes capable of infecting humans and aquatic animals, and the fact that V. vulnificus contains pathogenic as well as non-pathogenic variants. Here we report an alternative approach utilizing the development of a real-time PCR assay…
Development of a real-time PCR assay for detection and quantification of enterotoxigenic members of Bacillus cereus group in food samples
2009
A highly sensitive real-time PCR (qPCR) procedure, targeting the phosphatidylcholine-specific phospholipase C gene (pc-plc), was developed for specific detection and quantification of strains belonging to Bacillus cereus group. The target region was selected based on the enterotoxigenic profiles of 75 Bacillus strains. The inclusivity and exclusivity of the RTi-PCR assay were assessed with 59 isolates of the B. cereus group, 16 other Bacillus spp., and 4 non-Bacillus strains. The assay was also used to construct calibration curves for different food matrices, and it had a wide quantification range of 6 log units using both serial dilutions of purified DNA and calibrated cell suspensions of …
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%. …
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…
Direct Peel Monitoring of Xenobiotics in Fruit by Direct Analysis in Real Time Coupled to a Linear Quadrupole Ion Trap–Orbitrap Mass Spectrometer
2013
Study of xenobiotics present in fruit peel by exposing it (without any pretreatment) to direct analysis in real time coupled to a high-resolution orbitrap mass spectrometer (DART-HRMS) is reported for the first time. Variables such as DART gas heater temperature and pressure, source-to-MS distance, and sample velocity are investigated. The analysis of one sample by DART-MS lasts ca. 1 min, and the benefits of both high-resolution and tandem mass spectrometry to elucidate nontarget or unknown compounds are combined. Identification of postharvest fungicides, antioxidants, and sugars in fruit peel is performed in the positive ion mode. A possible elemental formula is suggested for marker compo…