Dual-channel biosensor for simultaneous detection of S. typhimurium and L. monocytogenes using nanotags of gold nanoparticles loaded metal-organic frameworks.

Simultaneous detection of multiple foodborne pathogens is of great importance for ensuring food safety. Herein, we present a sensitive dual-channel electrochemical biosensor based on copper metal organic frameworks (CuMOF) and lead metal organic framework (PbMOF) for simultaneous detection of Salmonella typhimurium (S. typhimurium) and Listeria monocytogenes (L. monocytogenes). The MOF-based nanotags were prepared by functionalizing gold nanoparticles loaded CuMOF (Au@CuMOF) and PbMOF (Au@PbMOF) with signal DNA sequences 1 (sDNA1) and sDNA2, respectively. By selecting invA of S. typhimurium and inlA gene of L. monocytogenes as targe sequences, a sandwich-typed dual-channel biosensor was developed on glassy carbon electrodes (GCE) through hybridization reactions. The sensitive detection of S. typhimurium and L. monocytogenes was achieved by the direct differential pulse voltametric (DPV) signals of Cu2+ and Pb2+. Under optimal conditions, channel 1 of the biosensor showed linear range for invA gene of S. typhimurium in 1 × 10-14-1 × 10-8 M with low detection limit (LOD) of 3.42 × 10-16 M (S/N = 3), and channel 2 of the biosensor showed linear range for inlA gene of L. monocytogenes in 1 × 10-13-1 × 10-8 M with LOD of 6.11 × 10-15 M (S/N = 3). The dual-channel biosensor showed good selectivity which were used to detect S. typhimurium with linear range of 5-1.0 × 104 CFU mL-1 (LOD of 2.33 CFU mL-1), and L. monocytogenes with linear range of 10 - 1.0 × 104 CFU mL-1 (LOD of 6.61 CFU mL-1).

An ultrasensitive biosensor for virulence ompA gene of Cronobacter sakazakii based on boron doped carbon quantum dots-AuNPs nanozyme and exonuclease III-assisted target-recycling strategy.

This work presented an electrochemical biosensor for the detection of virulence outer membrane protein A (ompA) gene of Cronobacter sakazakii (C. sakazakii), which was based on the mimic peroxidase activity of boron doped quantum dots-Au nanoparticles (BQDs-AuNPs) and a signal amplification strategy of exonuclease III (Exo III)-assisted target-recycling (EATR). The electrochemical signal was come from the electrochemical reduction of H2O2 by BQDs-AuNPs nanozyme. Due to the enhanced peroxidase-mimic electrocatalytic efficiency of BQDs-AuNPs and the EATR strategy, the biosensor showed a broad linear range (1.0 × 10-15 - 1.0 × 10-8 mol L-1) and a low limit of detection (LOD, 4.0 × 10-17 mol L-1). The constructed biosensor could also be applied in direct detection of extracted DNA from C. sakazakii. A good linear relationship (7.8 - 7.8 × 106 CFU mL-1) between the logarithm concentration of C. sakazakii and electrochemical signal was obtained with a LOD of 2.6 CFU mL-1. The biosensor was applied in the detection of impA gene segments in contaminated infant formula with recoveries ranged in 83.4 - 108.2%.

Electro-oxidation and determination 5-hydroxymethylfurfural in food on co-electrodeposited Cu-Ni bimetallic microparticles modified copper electrode.

This study presents a sensitive approach for electrochemical determination of 5-hydroxymethylfurfural (5-HMF) in food. The electrochemical sensor was fabricated on a copper electrode (CuE) modified with co-electrodeposited Cu-Ni bimetallic particles. This sensor, fabricated by 30 cycles of cyclic voltametric scanning with a scan rate of 50 mV s-1, exhibits good electrocatalytic ability to 5-HMF oxidation. Under the optimal conditions, linear scan voltammetry (LSV) and chronoamperometry were conducted for the determination of 5-HMF. The results of LSV show that a linear dependency within the 0.4-10 mM range with a detection limit (LOD) of 3.51 µM (S/N = 3) was achieved, while a linear range in 1 × 10-4-11 mM with a LOD of 0.043 µM (S/N = 3) was obtained by chronoamperometric measurement. The electrochemical sensor was finally applied in determination of 5-HMF in various foods, and the reliability and accuracy of the method were assessed by adopting an UV method as a standard method. Results show that the concentrations of 5-HMF in real samples are close to those measured by the standard method. In addition, standard addition method was further performed to evaluate the accuracy of our approach. The recoveries ranged from 90.0% to 110.0% are calculated, demonstrating good accuracy of the electrochemical sensor.

Colorimetric biosensing of nopaline synthase terminator using Fe3O4@Au and hemin-functionalized reduced graphene oxide.

In this paper, we present a simple and label-free colorimetric biosensor for detection of the nopaline synthase (NOS) terminator in genetically modified (GM) plants. The "signal on" colorimetric biosensor was developed using a nanocomposite consisted of gold nanoparticles doped magnetic Fe3O4 nanoparticles (Fe3O4@Au NP), capture probe DNA (cDNA), and hemin-functionalized reduced graphene oxide nanosheets (H-GN). The nanocomposite was successfully prepared by means of Au-S bonds and the strong π interactions between cDNA and H-GN. The sensing approach is based on the excellent peroxidase-mimicking activity of H-GN and its different electrostatic interactions with single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA). In presence of the target NOS, the cDNA in the nanocomposite will hybridize with its complementary sequence, and form dsDNA structure. Due to the weak π interactions between dsDNA and H-GN, a portion of H-GN will be released from the surface of Fe3O4@Au NPs and transferred into solution. After magnetic separation was performed, the supernatant was incubated with 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. The released H-GN can catalyze the oxidation reaction of TMB and turn the colorless solution blue. This "signal-on" colorimetric biosensor shows a broad linear range of 0.5-100 nM for the target NOS, with a 0.19 nM detection limit. The application of the biosensor for determination of NOS segments in samples of GM and non-GM tomatoes shows that it can discriminate between GM and non-GM plants. The reliability of the method for samples of NOS-spiked GM tomato suggests satisfactory recoveries in the range of 93.6%-94.2%.

Electrochemical detection of Salmonella using an invA genosensor on polypyrrole-reduced graphene oxide modified glassy carbon electrode and AuNPs-horseradish peroxidase-streptavidin as nanotag.

A rapid and sensitive electrochemical biosensor was constructed to detect Salmonella using invA gene biosensor. The biosensing was based on polyrrole-reduced graphene oxide (PPy-rGO) nanocomposite modified glassy carbon electrode (GCE) and signal amplification with horseradish peroxidase-streptavidin biofunctionalized gold nanoparticles (AuNPs-HRP-SA). PPy-rGO was prepared at 60 °C by chemical reduction of PPy-functionalized graphene oxide (PPy-GO) that was synthesized by in situ polymerization at room temperature. The detection signal was amplified via enzymatic reduction of H2O2 in the presence of hydroquinone (HQ) using AuNPs-HRP-SA as nanotag. Under optimal conditions, the differential pulse voltametric (DPV) signal from the biosensor was linearly related to the logarithm of target invA gene concentrations from 1.0 × 10-16 to 1.0 × 10-10 M, and the limit of detection (LOD) was 4.7 × 10-17 M. The biosensor can also detect Salmonella in the range of 9.6 to 9.6 × 104 CFU mL-1, with LOD of 8.07 CFU mL-1. The biosensor showed good regeneration ability, acceptable selectivity, repeatability and stability, which bode well as an alternative method for Salmonella screening.

[Moderate and severe persistent allergic rhinitis treated with acupoint application therapy of the different intensity: a randomized controlled trial].

OBJECTIVE: To evaluate the clinical therapeutic effects and safety on moderate and severe persistent allergic rhinitis treated with acupoint application therapy of the different intensity during the dog days. METHODS: One hundred and sixty patients of moderate and severe persistent allergic rhinitis were randomized into a No.1 treatment group, a No.2 treatment group, a No.3 treatment group and a placebo group, 40 cases in each one. The same acupoints were used in the four groups, named Dazhui (GV 14), Dingchuan (EX-B1), Feishu (BL 13), Pishu (BL 20), Mingmen (GV 4), Gaohuang (BL 43), Shenshu (BL 23) and Qihai (CV 6). In the three treatment groups, the fine powder of the ingredients (semen brassicae, radix angeliceae, asarum sieboldii, rhizome corydalis) of compound baijiezi formula was used. In the No.1 treatment group, the herbal paste (ginger-prepared paste) was prepared with ginger juice and the above herbal powder. In the No.2 and No.3 treatment groups, the herbal paste (honey-prepared paste) was prepared with honey with the above herbal powder. In the placebo group, the pseudo-herbal paste of the same appearance was prepared with millet powder and distilled water. The acupoint application was given for 2 h in the No.1 and No.2 groups and was for 6 h in the No.3 treatment group and the placebo group. The acupoint application therapy was given once every week during the dogdays, continuously for 5 weeks. The total nasal symptom score (TNSS), the score of the rhinoconjunctivitis quality of life questionnaire (RQLQ) and the count of blood eosinophils (EOS) were observed in the patients of the 4 groups before and after treatment. The clinical therapeutic effects were compared among the 4 groups. The incidences of the skin adverse reactions were observed in each treatment group. RESULTS: After treatment, the scores of TNSS and RQLQ were all reduced as compared with those before treatment in the three treatment groups (P<0.05, P<0.01), in which, the improvements in the No.3 treatment group were better than those in the No.1 treatment group and the No.2 treatment group (both P<0.05). After treatment, the count of EOS was all reduced as compared with that before treatment in the three treatment groups (all P<0.05). The differences were not significant statistically among the three treatment groups (all P>0.05). The total effective rate was 85.0% (34/40) in the No.3 treatment group, better than 76.3% (29/38) in the No.1 treatment group, 71.8% (28/39) in the No.2 treatment group and 5.0% (2/40) in the placebo group (P<0.05, P<0.01). The incidences of the skin adverse reaction in the No.3 treatment group and the No.2 treatment group were lower than those in the No.1 treatment group (both P<0.01). CONCLUSION: The acupoint application of the different intensity relieves the symptoms and improves the living quality in the patients of moderate and severe persistent allergic rhinitis. The stimulation of the ginger-prepared herbal paste is strong and induces skin blisters after 2 h herbal application. The stimulation of the honey-prepared herbal paste is moderate and does not induce blisters. The 6 h stimulation of the honey-prepared herbal paste is mild and the therapeutic effect is optimal.

Flow-injection determination of streptomycin residues in milk using the luminol-periodate-Mn2+ chemiluminescence system.

A novel flow-injection method with chemiluminescence detection was developed for the determination of streptomycin residues in milk, based on the enhancement by streptomycin of the luminol-potassium periodate-Mn(2+) chemiluminescence system. The assay allowed analysis of streptomycin residues in whole milk samples ( approximately 3.5% fat) after sample work-up procedures. The limit of detection was 5.16 x 10(-9) mol/L for milk, and was far below the regulations of maximum residue limits (MRLs). The relative standard deviation (RSD) for 8.66 x 10(-8) mol/L streptomycin was 1.94% (n = 15) in milk. The results obtained for the assay of streptomycin residues in milk was comparable with those obtained by the official methods (ELISA) and demonstrated good accuracy and precision. The possible mechanism is briefly discussed.

Determination of tetracyclines residues in honey using high-performance liquid chromatography with potassium permanganate-sodium sulfite-beta-cyclodextrin chemiluminescence detection.

A novel method was developed for the simultaneous determination of tetracycline antibiotic (TCA) residues such as oxytetracycline (OTC), tetracycline (TC), and metacycline (MTC) by high-performance liquid chromatography (HPLC) coupled with chemiluminescence (CL) detection. The procedure was based on the chemiluminescent enhancement by TCAs of the potassium permanganate-sodium sulfite-beta-cyclodextrin system in a phosphoric acid medium. The separation was carried out with an isocratic elution using a mixture of acetonitrile and 0.001 M phosphoric acid. For the three TCAs, the detection limits at a signal-to-noise of 3 ranged from 0.9 to 5.0 ng/ml. The relative standard deviations for the determination of TCAs ranged from 3.1 to 7.4% within a day (n=11) and ranged from 2.2 to 8.6% in 3 days (n=9), respectively. The method was successfully applied to the determination of TCA residues in honey samples. The possible mechanism of the CL reaction was also discussed.

Flow injection analysis of tetracyclines using inhibited Ru(bpy)3(2+)/tripropylamine electrochemiluminescence system.

Tetracyclines (TCs) were found to strongly inhibit the electrochemiluminescence (ECL) from the Ru(bpy)3(2+)-tripropylamine system when a working Pt electrode was maintained at 1.05 V (vs. Ag/AgCl) in pH 8.0 carbonate buffer solution. On this basis, a flow injection (FI) procedure with inhibited electrochemiluminescence detection has been developed for the determination of tetracycline (TC) and oxytetracycline (OTC). Under the optimized condition, the linear ranges of 2.0 x 10(-8)-1.0 x 10(-5) and 1.0 x 10(-8)-1.0 x 10(-5) g/mL and the detection limits of 4.0 x 10(-9) and 3.8 x 10(-9) g/mL were obtained for TC and OTC, respectively. The relative standard deviations (RSD) were 0.68% and 1.18% for 5.0 x 10(-7) g/mL TC and OTC (n = 13), respectively. The method showed higher sensitivity than most of the reported methods. It was successfully applied to the determination of tetracycline in a Chinese proprietary medicine, Tetracyclini and Cortisone Eye Ointment, and the residues of tetracycline in honey products. The inhibition mechanism has been proposed due to an energy transfer between electrogenerated Ru(bpy)3(2+)* and benzoquinone derivatives at the electrode surface.