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1.
Aim: To develop antibody–aptamer functionalized fibre‐optic biosensor for specific detection of Listeria monocytogenes from food products. Methods and Results: Aptamer, a single‐stranded oligonucleotide ligand that displays affinity for the target molecule, was used in the assay to provide sensor specificity. Aptamer‐A8, specific for internalin A, an invasin protein of L. monocytogenes, was used in the fibre‐optic sensor together with antibody in a sandwich format for detection of L. monocytogenes from food. Biotinylated polyclonal anti‐Listeria antibody, P66, was immobilized on streptavidin‐coated optical waveguide surface for capturing bacteria, and Alexa Fluor 647‐conjugated A8 was used as a reporter. The biosensor was able to selectively detect pathogenic Listeria in pure culture and in mixture with other bacteria at a concentration of approx. 103 CFU ml?1. This sensor also successfully detected L. monocytogenes cells from artificially contaminated (initial inoculation of 102 CFU 25 g?1) ready‐to‐eat meat products such as sliced beef, chicken and turkey after 18 h of enrichment. Conclusion: Based on the data presented in this study, the antibody–aptamer functionalized fibre‐optic biosensor could be used as a detection tool for sensitive and specific detection of L. monocytogenes from foods. Significance and Impact of the Study: The study demonstrates feasibility and novel application of aptamer on fibre‐optic biosensor platform for the sensitive detection of L. monocytogenes from food products.  相似文献   

2.
A novel flow injection analysis‐direct chemiluminescence (FI‐CL) method has been developed for determination of trace amounts of dopamine (DA) based on the enhancing effect of DA on the CL reaction of luminol with an Ag(III) complex in alkaline solution. Under optimum conditions, CL intensities are proportional to the concentration of DA in the range of 1.0 × 10?10 to 4.0 × 10?8 mol L?1. The detection limit is 3.0 × 10?11 mol L?1 for DA (3s), with a relative standard deviation (n = 13) of 2.3% for 1.0 × 10?8 mol L?1 DA. This method has also been applied for the determination of DA in commercial pharmaceutical injection samples. On the basis of the CL spectra and the results of the free‐radical trapping experiment of this work, a reaction mechanism for this CL reaction is proposed and discussed. Copyright © 2010 John Wiley & Sons, Ltd.  相似文献   

3.
Aims: To evaluate the effectiveness of the optimized immunomagnetic separation (IMS)‐plating protocol in relation to other culture, serological and molecular techniques currently used for Clavibacter michiganensis subsp. michiganensis in seed‐testing laboratories. Methods and results: Bacterial suspensions, tomato seed extracts spiked with the pathogen and naturally infected seeds were IMS‐plated for the detection of C. m. subsp. michiganensis. These results were compared with plating on general (YPGA) and semiselective (mSCM) media, double‐antibody sandwich enzyme‐linked immunosorbent assay (DAS‐ELISA), immunofluorescent assay (IF) or polymerase chain reaction (PCR). Different seed lots and pathogen strains were also tested. IMS‐plating allowed the detection of less than 10 CFU ml?1 of pathogen in all assayed samples. The mSCM medium provided positive results for 10 CFU ml?1 in naturally infected seeds, but up to 14 days was necessary for the typical colonies of the target to be come visible. By serological techniques, 103 and up to 104 CFU ml?1 were detected by IF and ELISA, respectively. DNA extraction was required to obtain positive results by PCR in seed extracts containing 103 CFU ml?1 or more. Conclusions: Among the evaluated methods, IMS‐plating provided the best results regarding sensitivity and specificity for C. m. subsp. michiganensis detection, allowing the recovery of viable bacteria from seed extracts. Significance and impact of the study: IMS‐plating increases isolation rates of C. m. subsp. michiganensis and could improve standard protocols currently used for routine analysis.  相似文献   

4.
Uniform molecular imprinting microspheres were prepared using precipitation polymerization with thifensulfuron‐methyl (TFM) as template, acrylamide as functional monomer and ethylene glycol dimethacrylate as cross‐linker. TFM could be selectively adsorbed on the molecularly imprinted polymers (MIPs) matrix through the hydrogen bonding interaction and the adsorbed TFM could be sensed by its strikingly enhancing effect on the weak chemiluminescence (CL) reaction between luminol and hydrogen peroxide. On this basis, a novel CL sensor for the determination of TFM using MIPs as recognition elements was established. The logarithm of net CL intensity (ΔI) is linearly proportional to the logarithm of TFM concentration (C) in the range from 1.0 × 10?9 to 5.0 × 10?5 mol L?1 with a detection limit of 8.3 × 10?10 mol L?1 (3σ). The results demonstrated that the MIP–CL sensor was reversible and reusable and that it could strikingly improve the selectivity and sensitivity of CL analysis. Furthermore, it is suggested that the CL enhancement of luminol–H2O2 by TFM might be ascribed to the enhancement effect of CO2, which came from TFM hydrolysis in basic medium. Copyright © 2010 John Wiley & Sons, Ltd.  相似文献   

5.
Qiyong Zhu 《Luminescence》2009,24(4):250-254
Based on the inhibition effect of rutin on the luminol–hydrogen peroxide chemiluminescence (CL) system catalyzed by tetrasulfonated colbalt phthalocyanine (CoTSPc), a sensitive flow‐injection CL method has been developed for the determination of rutin. The CL reaction mechanism was carefully investigated by examining CL emission spectra, UV–visible spectra and variation of reaction conditions. It was found that there existed a linear relationship between CL intensity and the concentration of rutin in the range of 8.0 × 10?9 to 1.0 × 10?6 mol L?1, and the detection limit is 3.8 × 10?9 mol L?1. This proposed method is sensitive, convenient and simple, and has been applied to the determination of rutin in commercial rutin tablets with satisfactory results. Copyright © 2009 John Wiley & Sons, Ltd.  相似文献   

6.
This article suggests a new sequential injection analysis chemiluminescence (SIA‐CL) strategy for monitoring the caffeine (CAF) content in soft and energy drinks using the catalytic activities of different nano‐metal oxides. The present study describes three different SIA‐CL systems (luminol–ferricyanide (III) coupled with Fe2O3 or ZnO nanoparticles (NPs), and luminol–H2O2 coupled with CuONPs. All experimental conditions were optimized and the linear concentration ranges of pure CAF were evaluated using the calibration graphs. The selectivity of the developed SIA‐CL systems was studied under the influence of various interfering species that may be present in soft or energy drinks such as sodium ions, sucrose, glucose, sodium benzoate, sodium citrate, riboflavin, niacin, citric, phosphoric and ascorbic acids. International Council for Harmonization (ICH) guidelines were obeyed for the validation of the suggested CL methods. The developed SIA‐CL systems displayed linear relationships over the concentration ranges 1.0–350, 5.0–400 and 10.0–400 μg ml?1 with Fe2O3 NPs, ZnO NPs and CuO NPs, respectively. The recorded lower limits of detection and quantification were 0.7, 2.7 and 7.8 μg ml?1, and 1.0, 5.0 and 10.0 μg ml?1 for the previously mentioned SIA‐CL systems. The results revealed high selectivity for CAF determination and were in good agreement with those obtained by other reported methods.  相似文献   

7.
Based on the catalytic activity of hemin, an efficient biocatalyst, an indirect capillary electrophoresis–chemiluminescence (CE‐CL) detection method for phenols using a hemin–luminol–hydrogen peroxide system was developed. Through a series of static injection experiments, hemin was found to perform best in a neutral solution rather than an acidic or alkaline medium. Although halide ions such as Br? and F? could further enhance the CL signal catalyzed by hemin, it is difficult to apply these conditions to this CE‐CL detection system because of the self‐polymerization of hemin, as it hinders the CE process. The addition of concentrated ammonium hydroxide to an aqueous/dimethyl sulfoxide solution of hemin–luminol afforded a stable CE‐CL baseline. The indirect CE‐CL detection of five phenols using this method gave the following limits of detections: 4.8 × 10?8 mol/L (o‐sec‐butylphenol), 4.9 × 10?8 mol/L (o‐cresol), 5.4 × 10?8 mol/L (m‐cresol), 5.3 × 10?8 mol/L (2,4‐dichlorophenol) and 7.1 × 10?8 mol/L (phenol). Copyright © 2013 John Wiley & Sons, Ltd.  相似文献   

8.
Hu Y  Li G  Zhang Z 《Luminescence》2011,26(5):313-318
In this paper, the novel trivalent copper–periodate complex {K5[Cu(HIO6)2], DPC} has been applied in a luminol‐based chemiluminescence (CL) reaction. Coupled with flow injection (FI) technology, the FI‐CL method was proposed for the determination of lincomycin hydrochloride. The CL reaction between luminol and DPC occurred in an alkaline medium. The CL intensity could be greatly enhanced by lincomycin hydrochloride. The relative CL intensity was proportional to the concentration of lincomycin hydrochloride in the range of 1 × 10?8 to 5 × 10?6 g mL?1 and the detection limit was at the 3.5 × 10?9 g mL?1 level. The relative standard deviation at 5 × 10?8 g mL?1 was 1.7% (n = 9). The sensitive method was successfully applied to the direct determination of lincomycin hydrochloride (ng mL?1) in serum. A possible mechanism of the lumonol–DPC CL reaction was discussed by the study of the CL kinetic characteristics and the spectra of CL reaction. The oxidability of DPC was studied by means of its electrochemical response. Copyright © 2010 John Wiley & Sons, Ltd.  相似文献   

9.
Based on the inhibition effect of methimazole (MMI) on the reaction of luminol–H2O2 catalyzed by gold nanoparticles, a novel chemiluminescence (CL) method was developed for the determination of MMI. Under the optimum conditions, the relative CL intensity was linearly related to MMI concentration in the range from 5.0 × 10?8 to 5.0 × 10?5 mol L?1. The detection limit was 1.6 × 10?8 mol L?1 (S/N = 3), and the RSD for 6.0 × 10?6 mol L?1 MMI was 4.83 (n = 11). This method has high sensitivity, wide linear range, inexpensive instrumentation and has been applied to detect MMI in pharmaceutical tablets and pig serum samples. Furthermore, a possible reaction mechanism is discussed. Copyright © 2010 John Wiley & Sons, Ltd.  相似文献   

10.
Aims: To investigate the suitability of human Hsp60, a receptor for Listeria adhesion protein (LAP), on paramagnetic beads (PMB) to capture Listeria monocytogenes from food in the presence of other Listeria to facilitate rapid and specific detection of this pathogen. Methods and Results: Commercially available streptavidin‐coated PMBs were linked with biotinylated Hsp60 (PMB‐Hsp60), and the bacterial capture efficiency from pure culture and meat samples was determined. Capture rate was also compared with the monoclonal antibody (MAb)‐C11E9‐coated beads (PMB‐C11E9) and the commercial Dynabeads anti‐Listeria. Captured cells were detected and quantified by plating on selective medium, quantitative real‐time PCR (qPCR) and a light‐scattering sensor. Overall, all ligand‐coated beads had similar capture efficiency (varied from 1·8 to 9·2%) for L. monocytogenes under the conditions employed, and the minimum cell number required to achieve such capture was 103 CFU ml?1. PMB‐Hsp60 had significantly greater capture efficiency for pathogenic Listeria (P < 0·0001) than the nonpathogenic Listeria. In contrast, PMB‐C11E9 and Dynabeads anti‐Listeria had similar capture efficiency for both. The efficacy of all PMBs to capture L. monocytogenes in the presence of Listeria innocua from food matrices was compared. Although Dynabeads anti‐Listeria had the overall best capture efficiency, PMB‐Hsp60 was able to selectively capture L. monocytogenes even in the presence of 10–100‐fold more L. innocua cells from enriched meat samples. Conclusions: Data show that the human cell receptor, Hsp60, is suitable for the capture of pathogenic Listeria on PMB in the presence of other Listeria in food. Significance and Impact of the Study: As pathogen interaction with host cells is highly specific, host cell receptors could be used as alternate capture molecules on PMB to aid in specific detection of pathogens.  相似文献   

11.
Chemiluminescence (CL) of the rhodamine 6‐G‐diperiodatonickelate (IV) (Rh6‐G‐Ni(IV) complex) in the presence of Brij‐35 was examined in an alkaline medium and implemented using flow‐injection analysis to analyze Mn(II) in natural waters. Brij‐35 was identified as the surfactant of choice that enhanced CL intensity by about 62% of the reaction. The calibration curves were linear in the range 1.7 × 10?3 – 0.2 (0.9990, n = 7) and 8.0 × 10?4 – 0.1 μg ml?1 (0.9990, n = 7) with limits of detection (LODs) (S:N = 3) of 5.0 × 10?4 and 2.4 × 10?4 μg ml?1 without and with using an in‐line 8‐hydroxyquinoline (8‐HQ) resin mini‐column, respectively. The sample throughput and relative standard deviation were 200 h?1 and 1.7–2.2% in the range studied respectively. Mn(II) concentrations in certified reference materials and natural water samples was successfully determined. A brief discussion about the possible CL reaction mechanism is also given. In addition, analysis of V(III), Cr(III) and Fe(II) was also performed without and with using an in‐line 8–HQ column and selective elution of each metal ion was achieved by adjusting the pH of the sample carrier stream with aqueous HCl solution.  相似文献   

12.
A simple and ultrasensitive flow injection chemiluminescence competitive immunoassay based on gold nanoparticle‐loaded enzyme for the detection of chloramphenicol (CAP) residues in shrimp and honey has been developed. Due to their good biocompatibility and large specific surface area, carboxylic resin beads can be used as solid phase carriers to immobilize more coating antigens (Ag). In addition, gold nanoparticles could provide an effective matrix for loading more CAP antibody and horseradish peroxidase, which would effectively catalyze the system of luminol–p‐iodophenol (PIP)–H2O2. A competitive immunoassay strategy was used for detection of CAP, in which CAP in the sample would compete with the coating Ag for the limited antibodies, leading to a chemiluminescence (CL) signal decrease with increase in CAP concentration. A wide linear range 0.001–10 ng ml?1 (R2 = 0.9961) was obtained under optimized conditions, and the detection limit (3σ) was calculated to be 0.33 pg ml?1. This method was also been successfully applied to determine CAP in shrimp and honey samples. The immunosensor proposed in this study not only has the advantages of high sensitivity, wider linear range, and satisfactory stability, but also expands the application of flow injection CL immunoassay in antibiotic detection.  相似文献   

13.
Aims: To determine if the purported deaminase inhibitors diphenyliodonium chloride (DIC) and thymol reduce the growth and survivability of Campylobacter. Methods and Results: Growth rates of Campylobacter jejuni and Camp. coli were reduced compared to unsupplemented controls during culture in Muellar–Hinton broth supplemented with 0·25 μmol DIC or thymol ml?1 but not with 0·01 μmol monensin ml?1 or 1% ethanol. Recovery of Camp. jejuni and Camp. coli was reduced >5 log10 CFU from controls after 24 h pure culture in Bolton broth supplemented with 0·25 or 1·0 μmol DIC ml?1 or with 1·0 μmol thymol ml?1. Similarly, each test Campylobacter strain was reduced >3 log10 CFU from controls after 24 h mixed culture with porcine faecal microbes in Bolton broth supplemented with 0·25 or 1·0 μmol DIC ml?1 or with 1·0 μmol thymol ml?1. Treatments with 0·25 μmol thymol ml?1, 0·01 μmol monensin ml?1 or 1% ethanol were less effective. Ammonia production during culture or incubation of cell lysates was reduced by 0·25 or 1·0 μmol DIC ml?1 but only intermittently reduced, if at all, by the other treatments. Conclusions: Diphenyliodonium chloride and thymol reduced growth, survivability and ammonia production of Camp. jejuni and Camp. coli. Significance and Impact of the Study: Results suggest a potential physiological characteristic that may be exploited to develop interventions.  相似文献   

14.
A novel chemiluminescence (CL) method was developed for the determination of cefazolin sodium based on the CL reaction between the [Cu(HIO6)2]5‐Cu(III) complex and luminol in alkaline solution. Results showed that CL emission of Cu(III) complex–luminol in alkaline medium was significantly different from that in acidic medium. A possible mechanism of the enhanced effect of cefazolin on CL emission of the [Cu(HIO6)2]5‐‐ luminol system was proposed. The effect of the reaction conditions on CL emissions was examined. Under optimized conditions, a good linear relationship was obtained between CL intensity and concentrations of cefazolin sodium in the range of 2.0 x 10‐8 to 2.0 x 10‐6 g/mL with a correlation coefficient of R2 = 0.9978. The limit of detection was 4.58 x 10‐9 g/mL. The proposed method was applied for the determination of cefazolin sodium in real samples with recoveries of 82.0‐109% with an RSD of 0.7‐2.1%. The proposed method was successfully used for the determination of cefazolin sodium in injectable powder preparations and human urine with satisfactory results. Copyright © 2012 John Wiley & Sons, Ltd.  相似文献   

15.
A simple and sensitive flow‐injection (FI) method for the determination of nitrate and nitrite in natural waters, based on luminol chemiluminescence (CL) detection, is reported. Nitrate was reduced online to nitrite via a copperized cadmium (Cu–Cd) column and then reacted with acidic hydrogen peroxide to form peroxynitrous acid. CL emission was observed from the oxidation of luminol in an alkaline medium in the presence of the peroxynitrite anion. The limits of detection (S:N = 3) were 0.02 and 0.01 µg N/L, with sample throughputs of 40 and 90 /h for nitrate and nitrite, respectively. Calibration graphs were linear over the range 0.02–50 and 0.01–50 µg N/L [R2 = 0.9984 (n = 8) and R2 = 0.9965 (n = 7)] for nitrate and nitrite, respectively, with relative standard deviations (RSDs; n = 3) in the range 1.8–4.6%. The key chemical and physical variables (reagent concentrations, buffer pH, flow rates, sample volume, Cu–Cd reductor column length) were optimized and potential interferences investigated. The effect of cations [Ca(II), Mg(II), Co(II), Fe(II) and Cu(II)] was masked online with EDTA. Common anions (PO43?, SO42? and HCO3?) did not interfere at their maximum admissible concentrations in freshwaters. The effect of salinity on the luminol CL reaction with and without nitrate and nitrite (2 and 0.5 µg N/L, respectively) was also investigated. The method was successfully applied to freshwaters and the results obtained were in good agreement with those obtained by an automated segmented flow analyser reference method. Copyright © 2011 John Wiley & Sons, Ltd.  相似文献   

16.
Tanshinol borneol ester (DBZ), a chemical combination of danshensu and borneol, is an experimental drug that exhibits efficacious anti‐ischemic activity in animal models. In this work, an ultrasensitive chemiluminescence (CL) method for the determination of DBZ was established based on the inhibitory effect of DBZ on the CL signal produced from the reaction between potassium permanganate and luminol in alkaline solution. The CL intensity responded linearly to the concentration of DBZ in the range 2.0 × 10‐10 to 4.0 × 10‐8 g/mL with a detection limit of 7 × 10?11 g/mL. The relative standard deviation (RSD) was 3.8% for 4.0 × 10?9 g DBZ (n = 11). The proposed method showed characteristics of high sensitivity, simple device and quick. In addition, this proposed method had been applied satisfactorily to the analysis of DBZ in blood. The pharmacokinetics of DBZ in rat has also been studied using the CL method. Copyright © 2013 John Wiley & Sons, Ltd.  相似文献   

17.
Aims: The microbial quality of farm bulk‐tank raw milk produced in Estonia during years 2004–2007 was investigated. Methods and Results: Bulk‐tank milk samples were analysed for lactic acid bacteria count (LABC), psychrotrophic bacteria count (PBC), aerobic spore‐forming bacteria count (ASFBC), total bacterial counts using BactoScan and somatic cell count (SCC) using Fossomatic. Randomly selected psychrotrophic isolates were subjected to 16S–23S PCR‐ribotyping. LABC remained below 104 CFU ml?1 in most samples, while psychrotrophic micro‐organisms dominated in 60% of farms. PBC ranged from 4·2 × 102 to 6·4 × 104 CFU ml?1, and ASFBC varied from 5 to 836 CFU ml?1. Conclusions: In general, the microbiological quality of the farm bulk‐tank milk was good – more than 91% of samples contained <50 000 CFU ml?1, and SCC in the majority of samples did not exceed the internationally recommended limits. Genus Pseudomonas spp. was the dominating spoilage flora with Pseudomonas fluorescens as the prevailing species. Significance and Impact of the Study: Specific bacterial groups (LABC, PBC and ASFBC), not analysed routinely by dairies, were determined in bulk‐tank raw milk of numerous dairy farms during 4‐year period. Based on the survey, dairy plants can better control their supply chains and select farms (milk) for the production of specific products, i.e. milk with low PBC and high LABC for cheesemaking.  相似文献   

18.
Aims:  Zero‐valent iron (ZVI) filters may provide an efficient method to mitigate the contamination of produce crops through irrigation water. Methods:  A field‐scale system was utilized to evaluate the effectiveness of a biosand filter (S), a biosand filter with ZVI incorporated (ZVI) and a control (C, no treatment) in decontaminating irrigation water. An inoculum of c. 8·5 log CFU 100 ml?1 of Escherichia coli O157:H12 was introduced to all three column treatments in 20‐l doses. Filtered waters were subsequently overhead irrigated to ‘Tyee’ spinach plants. Water, spinach plant and soil samples were obtained on days 0, 1, 4, 6, 8, 10, 13 and 15 and analysed for E. coli O157:H12 populations. Results:  ZVI filters inactivated c. 6 log CFU 100 ml?1E. coli O157:H12 during filtration on day 0, significantly (P < 0·05) more than S filter (0·49 CFU 100 ml?1) when compared to control on day 0 (8·3 log CFU 100 ml?1). On day 0, spinach plants irrigated with ZVI‐filtered water had significantly lower E. coli O157 counts (0·13 log CFU g?1) than spinach irrigated with either S‐filtered (4·37 log CFU g?1) or control (5·23 log CFU g?1) water. Soils irrigated with ZVI‐filtered water contained E. coli O157:H12 populations below the detection limit (2 log CFU g?1), while those irrigated with S‐filtered water (3·56 log CFU g?1) were significantly lower than those irrigated with control (4·64 log CFU g?1). Conclusions:  ZVI biosand filters were more effective in reducing E. coli O157:H12 populations in irrigation water than sand filters. Significance and Impact of the Study:  Zero‐valent ion treatment may be a cost‐effective mitigation step to help small farmers reduce risk of foodborne E. coli infections associated with contamination of leafy greens.  相似文献   

19.
A new chemiluminescence (CL) reaction between luminol and diperiodatoargentate {K2 [Ag (H2IO6) (OH) 2]} was observed in alkaline medium. The CL intensity could be greatly enhanced by amikacin sulfate. Therefore a new CL method for the determination of amikacin sulfate was built by combining with flow injection technology. A possible mechanism of the CL reaction was proposed via the investigation of the CL kinetic characteristics, the CL spectrum and the UV absorption spectra of some related substance. The concentration range of linear response was 5.1 × 10?8 to 5.1 × 10?6 mol L?1 with a detection limit of 1.9 × 10?8 mol L?1 (3σ). The proposed method had good reproducibility with a relative standard deviation of 2.8% (n = 7) for 5.1 × 10?7 mol L?1 of amikacin sulfate. It was successfully applied to determine amikacin sulfate in serum. Copyright © 2009 John Wiley & Sons, Ltd.  相似文献   

20.
A post‐chemiluminescence (PCL) phenomenon was observed when chloramphenicol was injected into a mixture of luminol and potassium periodate after the chemiluminescence (CL) reaction of luminol–potassium periodate had finished. The possible reaction mechanism was proposed based on studies of the CL kinetic characteristics, the CL spectra, the fluorescence spectra and the UV‐vis absorption spectra of the related substances. Based on the PCL reaction, a rapid and sensitive method for the determination of chloramphenicol was established. The linear response range was 6.0 × 10?7–1.0 × 10?5 mol/L, with a correlation coefficient of 0.9986. The relative standard deviation (RSD) for 5.0 × 10?6 mol/L chloramphenicol was 2.3% (n = 11). The detection limit was 1.6 × 10?7 mol/L. The method has been applied to the determination of chloramphenicol in pharmaceutical samples with satisfactory results. Copyright © 2011 John Wiley & Sons, Ltd.  相似文献   

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