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1.
The efficiency of 2 commercial enzyme-linked immunosorbent assay (ELISA) kits (Listeria-Tek and Tecra) for detecting Listeria in naturally contaminated foods was evaluated and compared with that of the culture method described in the Bacteriological Analytical Manual (BAM). Both ELISAs use modified University of Vermont (UVM-1) medium as a primary enrichment; the BAM method uses Listeria enrichment broth. Secondary enrichments for Listeria-Tek and Tecra, respectively, were Fraser broth and UVM-2, which contains additional acriflavin-HCl. When ELISA test results differed, secondary enrichments were tested against the other ELISA; Fraser broth was used to determine recovery rates because of its superiority over UVM-2. Of the 178 food samples examined, the presence of Listeria was detected and culturally confirmed in 38, 37, and 40 samples by the BAM, Listeria-Tek, and Tecra methods, respectively. Differences in results of the ELISAs compared with those of the BAM method were not statistically significant; however, differences between results of the 2 ELISA methods were significant. It was concluded that as rapid screening methods, the Listeria-Tek and the Tecra kits qualify as alternative methods to the BAM cultural method.  相似文献   

2.
Methodology for isolation of Listeria from foods--a Canadian perspective   总被引：4，自引：0，他引：4  
A previously described monoclonal antibody-enzyme immunoassay system for dairy products was examined to determine its potential for detecting Listeria in naturally contaminated ground meats. In addition, a microtiter plate system was developed which has potential in the rapid detection of Listeria species in foods. Experience in Canada with the U.S. Food and Drug Administration procedure for dairy products, the cold enrichment procedure, and the U.S. Department of Agriculture procedure for meats is discussed. Also, the status on attempts to devise improved selective media for the isolation of Listeria species from foods is described.  相似文献   

3.
A poultry rapid overnight field identification test (PROFIT) has been developed as a screening test which is practical, economical, and easy to perform and interpret for use in field environments to determine the presence of poultry tissue (chicken and turkey) in raw whole tissue or ground/formulated meat products. The basis of the test is an agar-gel immunodiffusion technique used with a printed template pattern and stabilized reagent paper discs. The test shows adequate sensitivity and specificity for its intended purpose. Key components are stable for at least 1 year if they are stored at refrigerator conditions. The design of the test is such that it can be made commercially available as a complete, stable, test kit suitable for use by any type of inspection program concerned with verification of poultry species in meat and/or poultry products that are subject to regulatory or quality controls.  相似文献   

4.
Historical perspectives on methodology to detect Listeria monocytogenes   总被引：6，自引：0，他引：6  
While recognized as a causative agent of illness in animals and humans for some time, the foodborne role of Listeria monocytogenes is a new and emerging one. This review briefly summarizes the historical developments in methodology used to detect the presence of L. monocytogenes. Although clinical procedures exist, these procedures do not consider isolation of Listeria from heavily contaminated environments. Federal agencies such as the U.S. Food and Drug Administration and the U.S. Department of Agriculture have defined protocols for the isolation of Listeria from dairy and meat products, respectively. Each of these protocols, and current problems common to all methods for the isolation of Listeria from food products, are discussed. Finally, future challenges with respect to improvement in our abilities to recognize, isolate, and rapidly identify Listeria in foods are presented.  相似文献   

5.
Two competitive immunoassays, a laboratory assay based on microwell plates and a field test based on the use of polystyrene tubes, have been developed for the detection of endosulfan in agricultural products. The limit of detection for the microwell plate format was 0.8 +/- 0.1 microg/kg, and the limit of detection for the tube format was 1.6 +/- 0.2 microg/kg. A simple, rapid, and efficient extraction method was employed, and 76-112% recoveries of spiked samples were obtained. Methanol extracts of some agricultural product samples such as grape, carrot, spinach, and tobacco could be analyzed directly by immunoassay after dilution in 0.5% fish skin gelatin-phosphate buffered saline. In contrast, extracts of green tea caused significant interference in the assay, and a number of simple cleanup methods were ineffective in removing interference. However, use of the coagulating reagent polyvinyl pyrrolidone removed the matrix effect effectively. For the validation of the enzyme-linked immunosorbent assay (ELISA) tests, samples were analyzed by ELISA and gas chromatography (GC) after solid phase extraction. The relationship between data obtained using the tube assay and microwell assay was good (the lowest r(2) value was 0.94), and also, the immunoassay assay data correlated well with data obtained from GC analysis (the lowest r(2) value was 0.93). The developed immunoassay methods are the suitable methods for the rapid quantitative and reliable determination of endosulfan residues in agricultural products.  相似文献   

6.
Background, Aim and Scope   A number of biotests are available for the characterisation of solid matters such as soil or sediment. Among these, bacterial biotests using single test species often analyse the toxicity of water-soluble contaminants in aquatic extracts, but there is also a need for a fast and inexpensive bacterial solid-contact test. In this study, a solid contact test with added bacteria (Arthrobacter globiformis) was optimised (through miniaturization) for the development of a test kit with conserved bacteria. As in other tests, the results can be influenced by natural soil factors, often masking anthropogenic impacts. For this reason, a further goal of this study was the investigation of the influence of natural soil characteristics on the result of the solid contact test. The project is part of the joint research project 'Optimization of ecotoxicological test methods for routine use' (abbreviated as ERNTE-Forschungsvorhaben: 0330305). Materials and Methods: This method is based on an existing German standard (DIN 38412 L 48) using Arthrobacter globiformi for testing whole soils and sediments. The test principle is the measurement of the dehydrogenase activity of the test organism A. globiformis after an incubation time of two hours with the solid material. To attain the miniaturization in microplates, dye measurement was changed from spectrophotometrical determination of the substrate resazurine to the fluorimetric measurement of the product resorufin. A second step towards optimisation was the use of freeze-dried bacteria. Seven selected uncontaminated soils were tested in order to determine the influence of natural soil characteristics on the results of the solid contact test with A. globiformis. Freshly spiked and polluted field soils were analysed in order to obtain information about the sensitivity of the test. Results: It is possible to perform the contact test in microplates. The fluorimetric dye measurement can be carried out in the presence of the solid material, so the work-intensive step of centrifugation and filtration is no longer necessary. The measurement in the optimised contact test is based on the kinetics of the enzyme reaction. The investigation showed that conserved bacteria have the same activity and sensitivity as cultivated bacteria. Discussion: The study of the uncontaminated soils demonstrated the influence of various soil characteristics on the results of the solid contact test. This information is the basis for the selection of the control and reference soils and is crucial for setting the threshold value in toxicity testing. The investigation of freshly spiked and contaminated soils showed a different sensitivity dependent on the kind of the contamination. Conclusions: The solid contact test was successfully optimised using microplates, whereas now less than six hours are necessary for the analysis. The optimised test is rapid and sensitive, requiring small samples and no stock culture of the bacteria A. globiformis if using freeze-dried bacteria. In this study, the effect of natural soil factors such as pH-value was shown. This information is used to define the threshold value for toxicity. Therefore, the optimised contact test can be used for an efficient assessment of soil or soil substrates. Further studies will clear up if this optimisation is also valid for aquatic sediments and waste. Recommendations and Perspectives: Due to its short analysis time, the test is suitable for screening different kinds of solid matter and can be used for on-site analysis. - The optimised contact test with freeze-dried bacteria as part of a battery of tests is appropriate for the assessment of contaminated soils, sediments and waste.  相似文献   

7.
A wide variety of enzyme‐linked immunosorbent assays (ELISAs) are commercially available for gluten detection in food, including new formats and assays with antibodies against relevant gluten epitopes. Nevertheless, problems persist to accurately determine the gluten content of products. In this study, the performance of a set of 14 ELISA kits for gluten detection, representative of the current ELISA methods available on the market, was evaluated. These tests were used to determine gluten content in a series of relevant food matrices varying in complexity. Our results show that, currently, there is no single ELISA method that can accurately detect and quantify gluten in all different matrices. This includes the current type I method R5 as recommended by Codex Alimentarius. We conclude that further improvements are urgently needed and recommend focusing on competitive formats, improving extraction methods, and the detection of relevant gluten peptides (in order of priority).  相似文献   

8.
A total of 1074 test samples of commercial, domestic, vacuum-packaged fresh fish were studied to determine whether spoilage occurs before the products become toxic from naturally occurring Clostridium botulinum spores. The products were incubated for 12 days at 12 degrees C (mild abuse). After incubation, they were tested for botulinal toxin and evaluated for organoleptic acceptability. Even when only marginally acceptable to laboratory personnel, none of the 1074 test samples were positive for C. botulinum toxin. Thus, the fish either contained no C. botulinum spores, or the spores were unable to grow out and produce toxin before spoilage made the product marginally unacceptable.  相似文献   

9.
Abstract

Nitrate remains a contaminant of concern for users of well water. Well‐water evaluation, either to assess nitrate contamination or to evaluate sites prior to including them in a larger water quality study, often involves costly laboratory analysis. A cost effective alternative to laboratory analysis are dip‐style test strips. However, the accuracy of these types of products must be reliable, as failure to identify the contaminant may, for example, persuade a homeowner to neglect to have a potential problem further investigated. The testing of nitrate using such strips typically involves dipping the strip into the water sample and reading the color development after a specific period of time. The color development is then compared to a color scale which corresponds to concentration provided with the test. The results of these types of analysis are especially open to interpretation by the evaluator of the results. An experiment was conducted to evaluate test strips in which individuals or “readers” tested water samples collected in the field and nitrate standards prepared in the laboratory with nitrate test strips. The results obtained by the “readers” were compared to analysis of nitrate by high performance liquid chromatography and colorimetric analysis using a colorimetric ion analyzer. There was a good agreement between the “readers” results and the analytical methodologies used. Use of the test strips by non‐technical persons, such as homeowners, could provide an accurate determination of nitrate in well water without the expense involved in a detailed laboratory analysis. The test strips can also be relied upon to accurately determine nitrate concentration when screening wells prior to designing field experiments.  相似文献   

10.
Listeria monocytogenes, one of the "new enemies" in food microbiology, is a human and animal pathogen that is widespread in nature. The organism is a transient constituent of the intestinal flora excreted by 1-10% of healthy humans. It is an extremely hardy organism and can survive for many years in the cold in naturally infected sources. L. monocytogenes has been isolated from a wide variety of foods, including dairy products, meats, and fish. Although most of the foodborne listeriosis outbreaks have been linked to the consumption of dairy products, recent sporadic cases have been associated with meats, as well as other foods. It is now recognized that listeriolysin 0, a 60-kilodalton protein, is one of the major virulence factors of the organism. All strains of L. monocytogenes are pathogenic by definition although some appear to be more virulent than others. There have been recent reports of hemolytic isolates of L. monocytogenes, which are nonpathogenic for mice. Attachment to and penetration of cells also appear to be prerequisites for human infection. Cultural methodology for isolating the organism from foods has been in a state of flux since 1985. Rapid methods using both ELISA and DNA technology have been developed. Because of the widespread nature of the organism, it is nearly impossible to eliminate it from the food supply. However, by using a hazard analysis-critical control point approach, the health hazard associated with this organism can be reduced to a minimum.  相似文献   

11.
Hidden allergens in food products are, especially for peanut-allergic consumers, a serious problem because even low amounts (approximately 200 microg) of peanut can elicit allergic reactions. Undeclared peanut traces can be found in processed food products, because contaminations with peanut during production processes are frequent. To minimize the risk of such cross-contaminations, it is necessary to develop sensitive analytical methods for the detection of hidden allergens in foods. For this approach we developed two peanut-specific assays based on the detection of peanut protein by specific antibodies (sandwich ELISA) and by the detection of peanut-specific DNA (part of the coding region of Ara h 2) by a real-time PCR. Both tests did not show any cross-reactivity with 22 common food ingredients (cereals, nuts, legumes), and the limit of detection is <10 ppm peanut in processed foods. Thirty-three random samples of food products were tested for the presence of peanut to compare both assay types with each other and to evaluate the percentage of foods on the German market that are contaminated with peanut traces. We found that four products (13.3%) without peanut in the list of ingredients contained peanut protein in a range from 1 to 74 ppm peanut protein and that the results of both tests correlated well. The real-time PCR was able to detect one more positive sample than the sandwich ELISA. In conclusion, both assays are sensitive and specific tools for the detection of hidden allergens in processed foods.  相似文献   

12.
研究了芹菜花生冷食菜的辐照杀菌效果和接种于其中的致病菌(Listeria innocua L83和Salmonellaenteritidis500041)的辐照效应,以及辐照处理对芹菜花生冷食菜营养品质和感官品质的影响。结果表明,2kGy辐照处理能有效降低冷食菜中微生物的含量,辐照接种于其中的沙门氏菌和李斯特菌的D10值分别为0.284和0.296 kGy,2.0kGy以下的辐照剂量对其感官品质和蛋白质的营养品质没有明显的影响。2.0kGy剂量能使冷食菜中的微生物降低10-2~10-3,使接种于其中的致病菌降低10-6以上,能有效地提高芹菜花生冷食菜的卫生安全。  相似文献   

13.
A sandwich enzyme-linked immunosorbent assay (ELISA) has been developed for detection of α-amylase in preharvest sprouted wheat and adapted to rapid field-use formats requiring 15–20 min to perform. Polyclonal and monoclonal antibodies were prepared to detect a mixture of high and low pI isozymes of α-amylase and high pI isozymes only. All antibodies detected α-amylase on immunoblots of either a crude wheat extract or of purified enzyme, but only the polyclonal antibodies functioned in a sandwich ELISA. Depending on the antibody combination, the tube ELISA detected either the high and low pI isozymes of α-amylase or the high pI isozymes only with a detection limit of ≈0.5–1.0 ng/mL of amylase. Wheats with falling numbers (FN) of <350 sec could be discriminated from sound wheats, with decreasing FN producing increasing assay color. Using 130 wheat grain samples, ELISA absorbances for detection of both high and low pI isozymes and of high pI isozymes only were highly positively correlated with amylase enzyme activity and negatively correlated with FN. The correlations were similar for detection of both isozyme families and for detection of high pI isozymes only. Analyses of three sets of wheat samples from different environments demonstrated that the relationship between ELISA absorbance and FN had little dependence on wheat cultivar. The precision of sample analysis using the field ELISA was similar to the precision of FN test apparatus.  相似文献   

14.
A corroborative study was conducted on the maize quality properties of test weight, pycnometer density, tangential abrasive dehulling device (TADD), time-to-grind on the Stenvert hardness tester (SHT), 100-kernel weight, kernel size distribution, and proximate composition as well as maize dry- and wet-millability by six participating laboratories. Suggested operating procedures were given to compare their measurements and provide the variance structure within and between laboratories and hybrids. Partial correlation coefficient among maize quality properties varied among laboratories. The repeatability and reproducibility precision values were acceptably low for the physical quality tests, except for TADD and SHT time-to-grind measurements. The yields of dry- and wet-milled products and their correlation with maize quality properties were dependent on the collaborating laboratory. This paper highlights the importance of laboratory variation when considering which maize hybrids are best suited for dry-milling and wet-milling.  相似文献   

15.
A porcine rapid identification method (PRIME) has been developed for detection of pork in a wide variety of meat products. The test is an adaptation of previously developed field screening immunodiffusion tests for beef and poultry detection. The PRIME test was demonstrated to be specific, sensitive, and accurate in the analysis of samples in the laboratory and in a commercial meat processing establishment.  相似文献   

16.
We evaluated a new method to measure in situ denitrification under field conditions in a number of water-saturated subsoils that had a broad range of biogeochemical properties. A test solution containing 15NO3− and/or C2H2 was introduced to the subsoil and the subsequent production of dissolved denitrification products was measured to quantify denitrification activity. The method showed a clear production of denitrification products over time. Results were compared to laboratory-based measurements from the same soil incubated as anaerobic slurries with added 15NO3−. Rates of denitrification with the in situ and the laboratory methods ranged from 1-2800 and 1-1700 μg N kg−1 d−1, respectively. Generally the methods gave good agreement and we consider both to be valid. However, there were some significant deviations, which we attribute to spatial heterogeneity and laboratory effects. Because the laboratory method is so much easier to perform, we suggest it should be the preferred method for large-scale studies of denitrification from the soil types we investigated. However, the two methods showed poor agreement in determining the proportion of N2O in the total denitrification output. This was because this proportion is subject to delicate and complex control. We conclude that neither method was suitable for quantifying N2O emission from the denitrification measurements.  相似文献   

17.
The development of an enzyme-linked immunosorbent assay (ELISA) based on polyclonal antibodies for a class of endocrine disrupting compounds, 4-nonylphenol, is described. The parent molecule was derivatized at the ortho position of the free phenolic hydroxyl group to obtain the hapten, NP1, and it was conjugated with keyhole limpet hemocyanin, which was used as an immunogen. Four antisera were generated and screened against three coating antigens. The most sensitive ELISA from the screening tests (antiserum NP03As, 1/1000, and coating antigen NP1-BSA, 1 microg/mL) was further optimized and characterized. The influence of various physicochemical factors (organic solvent, pH, ion strength) was investigated. Methanol as the additive organic solvent was found to be the best organic solvent for the ELISA, with optimal sensitivity observed at a concentration of 5%. The ELISA parameters were changed at more acidic or basic pH values, whereas higher ionic strengths strongly suppressed the I(50) value and the maximum absorbance. The most sensitive ELISA for 4-nonylphenol exhibited an I(50) value of 38.6 +/- 5.5 microg/L, with a dynamic range from 12 to 350 microg/L, and the lower limit of detection was 7.7 +/- 1.3 microg/L. The optimized ELISA displayed no significant cross-reaction against the parent compounds, nonylphenol ethoxylates, degradation products, carboxylates, and bisphenol A, except in 4-octylphenol.  相似文献   

18.
The feeding of ruminant proteins to ruminants is prohibited in most countries because the practice is thought to be responsible for the spread of bovine spongiform encephalopathy. However, currently available methods to detect ruminant blood products in rendered feedstuffs are inadequate because they lack species specificity, tissue specificity, and are not based on a thermostable analyte. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for this study that provides reliable and sensitive (0.05-0.5% v/v) detection of bovine blood materials in animal feed. The new sandwich ELISA employs two previously developed monoclonal antibodies (MAbs), Bb6G12 as the capture antibody and biotinylated MAb Bb3D6 as the detecting antibody, and is bovine-specific and blood-specific. The assay is based on the detection of a 60 kDa thermostable protein in bovine blood and provides a useful regulatory tool for monitoring fraudulent labeling or contamination of bovine blood in both heat-processed feedstuffs and unprocessed raw materials. Keywords: Bovine; blood; monoclonal antibody; sandwich ELISA.  相似文献   

19.
The nonenzymatic glycation of proteins by reducing sugars, also known as the Maillard reaction, has received increasing recognition from nutritional science and medical research. In this study, we applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to perform relative and simultaneous quantification of the Amadori product, which is an early glycation product, and of N(epsilon)-(carboxymethyl)lysine and imidazolone A, two important advanced glycation end products. Therefore, native lysozyme was incubated with d-glucose for increasing periods of time (1, 4, 8, and 16 weeks) in phosphate-buffered saline pH 7.8 at 50 degrees C. After enzymatic digestion with endoproteinase Glu-C, the N-terminal peptide fragment (m/z 838; amino acid sequence KVFGRCE) and the C-terminal peptide fragment (m/z 1202; amino acid sequence VQAWIRGCRL) were used for relative quantification of the three Maillard products. Amadori product, N(epsilon)-(carboxymethyl)lysine, and imidazolone A were the main glycation products formed under these conditions. Their formation was dependent on glucose concentration and reaction time. The kinetics were similar to those obtained by competitive ELISA, an established method for quantification of N(epsilon)-(carboxymethyl)lysine and imidazolone A. Inhibition experiments showed that coincubation with N(alpha)-acetylargine suppressed formation of imidazolone A but not of the Amadori product or N(epsilon)-(carboxymethyl)lysine. The presence of N(alpha)-acetyllysine resulted in the inhibition of lysine modifications but in higher concentrations of imidazolone A. o-Phenylenediamine decreased the yield of the Amadori product and completely inhibited the formation of N(epsilon)-(carboxymethyl)lysine and imidazolone A. MALDI-TOF-MS proved to be a new analytical tool for the simultaneous, relative quantification of specific products of the Maillard reaction. For the first time, kinetic data of defined products on specific sites of glycated protein could be measured. This characterizes MALDI-TOF-MS as a valuable method for monitoring the Maillard reaction in the course of food processing.  相似文献   

20.
A DNA probe was used to identify hemolytic Listeria monocytogenes in naturally contaminated dairy products: unpasteurized milk, ricotta cheese, and imported semisoft cheeses. Of 34 milk samples, 12 were suspected to contain hemolytic L. monocytogenes; 1 contained greater than 6000 viable organisms/g. The ricotta cheese, although temperature-abused, had a titer of 3.6 x 10(6) beta-hemolytic L. monocytogenes cells/g, whereas the semisoft cheeses reached a maximum of 5.6 x 10(6) cells/g. Pure cultures of L. monocytogenes isolated from both types of cheese were found positive by the CAMP test and the DNA probe.  相似文献   

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