Simultaneous and sensitive determination of Escherichia coli O157:H7 and Salmonella Typhimurium using evanescent wave dual-color fluorescence aptasensor based on micro/nano size effect.

The simultaneous and sensitive determination of two common pathogenic bacteria, Escherichia coli O157:H7 (E. coli O157:H7) and Salmonella Typhimurium (S. Typhimurium) was achieved using evanescent wave dual-color fluorescence aptasensor and the fiber nanoprobe through combining the micro/nano size and time-resolved effect. Two fluorescence labeled aptasensors, Cy3-apt-E and Cy5.5-apt-S, were regarded as biorecognition elements and signal reporters for E. coli O157:H7 and S. Typhimurium, which were alternatively excited by evanescent waves originated from 520 nm to 635 nm excitation lights, respectively. The fiber nanoprobe with in-situ etched nanopores was used for distinguishing free aptasensors and aptasensors bound to pathogenic bacteria based on the limited penetrated depth of evanescent wave and the significant size difference of bacteria and nanopore. The E. coli O157:H7 and S. Typhimurium were directly and simultaneously quantitated in less than 35 min without the requirement of the complex immobilization of biorecognition molecules and bacteria enrichment/separation processes. The limits of detection of E. coli O157:H7 and S. Typhimurium were 340 CFU/mL and 180 CFU/mL, respectively. The satisfied recovery rate of real samples testing verified the feasibility and accuracy of the proposed method. Our strategy not only greatly simplifies the detection and identification process of multiple pathogenic bacteria, but also is easy to extend as a universal technology for sensitive determination of other bacteria using their respective biorecognition molecules.

Portable and automated fluorescence microarray biosensing platform for on-site parallel detection and early-warning of multiple pollutants.

The portable and automated fluorescence microarray biosensing platform (FMB) that employed a compact hybrid optical structure, microfluidics, and microarray biosensors was constructed for on-site parallel detection of multiple analytes. In the FMB, a hybrid optical structure that composed of a 1 × 4 single mode fiber optic coupler, four fiber optic switches, a single-multi mode fiber optic bundle coupler was at the first time developed for the transmission of the excitation light and the collection and transmission of multi-channel fluorescence signals. Through the control of fiber optic switches, the parallel fluorescence assay of four channels could be achieved using only one excitation light and one photodiode detector on the basis of the time-resolved effect. This optical design not only greatly increased the efficiency of light transmission and fluorescence collection and detection sensitivity of the FMB, but also allows the miniaturization and portability of the whole system because of few optical separation elements used and no requirement of rigorous optical alignment. Taking Microcystin-LR (MC-LR), 2,4-D, atrazine (ATZ), and bisphenol A (BPA) for example, the application potential of the FMB to rapidly and parallelly detect four typical pollutants in real water with high sensitivity and specificity was demonstrated. The limits of detection of MC-LR, 2,4-D, ATZ, and BPA were 0.04 µg/L, 0.09 µg/L, 0.02 µg/L, and 0.03 µg/L, respectively. The FMB could also achieve early-warning of pollutants thanks to its ability of rapidity, high-frequency, and multiple-analyte detection. The FMB has significant implications as a multiplexable, portable, rapid, and quantitative detection platform for pollution accidents and water quality management to satisfy the increasing demands of alerting and protecting civilians.

A novel dual-color total internal reflection fluorescence detecting platform using compact optical structure and silicon-based photodetector.

A novel dual-color total internal reflection fluorescence detecting platform (DTP) was developed for the simultaneous detection of two fluorescence dyes. The DTP employed a simple and compact optical system and a Si-based photodetector SOP-1000 assembly to improve the optical efficiency and detection sensitivity. In the optical system, a single-multimode fiber optic coupler was applied for the transmission of two wavelength excitation lights and the collection and transmission of two wavelength fluorescence signals, and a fiber optical switch controlled two wavelength excited lights to alternatively enter into the combined tapered fiber optic probe and two wavelength fluorescence dyes were alternatively excited. The simultaneous and sensitive detection of two wavelength fluorescence signals was achieved by one photodetector SOP-1000 based on the time-resolved effect of the fiber optical switch. The DTP demonstrated its sensitivity of 50 fW light intensity, and the limit of detection of Cy5.5 and Pacific Blue dye (PB) were 0.05 nM and 2.1 nM, respectively. This miniaturized and integrated DTP with high sensitivity, simple optical structure, negligible cross-talk, and cost-effectiveness could be a potential alternative to the conventional dual-color fluorescence detecting systems. It could also be a powerful component of a µ-TAS system for highly sensitive dual-color fluorescence detection.

SERS based aptasensor for ochratoxin A by combining Fe3O4@Au magnetic nanoparticles and Au-DTNB@Ag nanoprobes with multiple signal enhancement.

A SERS-based aptasensor for ochratoxin A (OTA) is described. It is making use of Fe3O4@Au magnetic nanoparticles (MGNPs) and of Au@Ag nanoprobes modified with the Raman reporter 5,5-dithiobis-(2-nitrobenzoic acid; DTNB). Au-DTNB@Ag NPs were modified with the OTA aptamer (aptamer-GSNPs) and used as Raman signal probes. The SERS peak of DTNB at 1331 cm-1 was used for quantitative analysis. MGNPs modified with cDNA (cDNA-MGNPs) were used as capture probes and reinforced substrates. When the Au-DTNB@Ag-Fe3O4@Au complexes are formed through oligonucleotide hybridization, the Raman signal intensity of the Raman probe is significantly enhanced. If the OTA concentration in samples increases, more Raman signal probes (aptamer-GSNPs) will dissociate from the cDNA-MGNPs because more OTA aptamer is bound by OTA. This leads to a lower Raman signal after magnetic separation. Under the optimal conditions, the detection limit for OTA is 0.48 pg·mL-1 based on 3σ criterion. This is attributed to the multiple Raman signal enhancement and the good performance of the OTA aptamer. The good recovery and accuracy of the assay was confirmed by evaluating spiked samples of wine and coffee. Graphical abstract Schematic of an aptamer based SERS assay for OTA by integrating Fe3O4@AuNPs (MGNPs) with Au-DTNB@Ag NPs with multiple signal enhancement. Aptamer modified Au-DTNB@Ag NPs are used as Raman probes, and MGNPs modified with cDNA are used as capture probes and reinforced substrates.

A FRET-based dual-color evanescent wave optical fiber aptasensor for simultaneous fluorometric determination of aflatoxin M1 and ochratoxin A.

A dual-color fluorescence resonance energy transfer (FRET) based aptasensor is described for simultaneous determination of the mycotoxins aflatoxin M1 (AFM1) and ochratoxin A (OTA). Aptamers against AFM1 and OTA were labeled with two fluorophores with different excitation wavelengths (Cy5.5; 675 nm; and Alexa 405; 401 nm), respectively. They were used as the signalling probes. A compact dual-color evanescent wave all-fiber detection system with two lasers (635 nm; red; and 405 nm; purple) was used for the simultaneous collection of two-wavelength fluorescence signals. The hybridization of labeled aptamers with complementary sequences (Q-cDNA) labeled with a dark quencher (BHQ3 or dabcyl) causes fluorescence to be strongly reduced because of the fluorescence resonance energy transfer. In the presence of AFM1 and OTA, they bind to their respective aptamer and result in the dissociation of double stranded DNA, which induce fluorescence recovery. Under the optimum conditions, AFM1 and OTA can simultaneously and selectively be determined ranged from 1 ng·L-1 to 1 mg·L-1. The detection limits of AFM1 and OTA are 21 and 330 ng·L-1, respectively (S/N = 3). The FRET-based dual-color detection scheme was applied to the simultaneous detection of AFM1 and OTA in milk with good recovery, precision, and accuracy. Graphical abstract Aptamers against AFM1 and OTA were labeled with two fluorophores with different excitation wavelengths (Cy5.5; 675 nm; and Alexa 405; 401 nm) and then used as signalling probes. A FRET-based aptasensor is described for simultaneous determination of AFM1 and OTA using dual-color evanescent wave system with two lasers (635 nm; red; and 405 nm).

Development of novel portable and reusable fiber optical chemiluminescent biosensor and its application for sensitive detection of microcystin-LR.

A novel portable fiber optical chemiluminescent biosensor (FOCB) system was successfully constructed using fiber optic bio-probe as biorecognition element as well as transducer and a Si-based photodiode detector (PD-3000). The compact design of FOCB with free magnetic beads and photomultiplier allows it to be portable and suitable for on-site and automatic detection of targets, as well as to be cost-effective and small sample volume (only 40.0 µL). The sensitivity of 5 fW could be obtained for the FOCB due to the high performance of PD-3000 and no background light signal. Next, the FOCB was applied for the CL-based detection of microcystin-LR (MC-LR) in water using hapten-carrier protein conjugates modified fiber optic bio-probe as biorecognition element. Based on indirect competitive sandwich-like CL immunoassay principle, the highly sensitive detection of MC-LR was achieved using FOCB system. Under optimal conditions, the limit of detection for MC-LR is 0.03 µg/L, which is comparable with that of most reported MC-LR immunoassay methods. The robustness of the hapten-carrier protein-modified biosensor surface allowed multiple MC-LR immunoassays without any significant loss of performance, which ensured accurate and cost-effective detection results. The proposed strategy demonstrated good recovery, precision, and accuracy through the evaluation of the spiked water samples. The FOCB system can be readily extended toward the on-site real-time sensitive detection of other targets in the field of environment, food, and medical diagnosis.

Universal and reusable hapten/antibody-mediated portable optofluidic immunosensing platform for rapid on-site detection of pathogens.

A universal and reusable hapten-antibody-mediated portable optofluidic immunosensing platform (OIP) was developed for rapid on-site detection of pathogens. By using Escherichia coli O157:H7 (E. coli O157:H7) and bisphenol A-Bovine serum albumin (BPA-BSA)/anti-BPA antibody as a model pathogen and a mediated hapten-antibody, respectively, a novel immunoassay mechanism was proposed to detect pathogens. The BPA-BSA-modified immunosensor and E. coli O157:H7 were initially saturated with anti-BPA antibodies (mouse IgG) and anti-E. coli O157:H7 antibodies (mouse IgG), respectively. Then, the fluorescence-labeled secondary antibodies (goat anti-mouse IgG antibody) were incubated with E. coli O157:H7 with their antibodies. Next, the mixture was introduced into the immunosensor surface bound to the anti-BPA antibodies. A high concentration of E. coli O157:H7 in the sample reduced the number of fluorescence-labeled secondary antibodies bound to the immunosensor surface, thus resulting in the detection of low fluorescence signals. Under optimized conditions, the hapten-antibody-mediated OIP system exhibited a detection limit of 8 cfu/mL E. coli O157:H7 after concentrating 100 times by using centrifugation, and a test cycle, including prereaction, detection, and regeneration, was less than 1 h. The robustness of the hapten-carrier protein-modified immunosensor surface allowed multiple pathogen immunoassays. The proposed strategy demonstrated good recovery, precision, and accuracy through the evaluation of the spiked water samples. We expect that the new platform can be readily used for the detection of other pathogens in a variety of application fields ranging from environmental monitoring and food safety to medical diagnosis.

Effects and mechanism of diclofenac degradation in aqueous solution by US/Zn0.

A system of ultrasound radiation coupled with Zn0 was applied to degrade diclofenac. The effects of initial pH, dosage of Zn0 and ultrasound density were investigated. To further explore the mechanism of the microcosmic reaction, the fresh and used Zn0 powders were characterized by SEM, XRD and XPS. Radical scavengers were used to determine the oxidation performance of strong oxidizing free radicals on diclofenac, including hydroxyl radicals and superoxide radicals. The results showed that the optimum removal of diclofenac reached to over 85% at pH of 2.0 in 15min, with Zn0 dosage of 0.1g/L and ultrasound density of 0.6W/cm3. TOC removal of 72.6% in 15min and dechlorination efficiency of diclofenac reached 70% in 30min. Characterization results showed that a ZnO membrane was generated on the surface of Zn particles after use. According to the mass spectrometry results, several possible pathways of diclofenac degradation were proposed, and most diclofenac was turned into micro-molecules or CO2 finally. The synergistic effect of US/Zn0 in the reactions led to a proposed degradation mechanism in which zinc could directly attack the target contaminant diclofenac because of its good reducibility with the auxiliary functions of ultrasonic irradiation, mechanical shearing and free radical oxidation.

NiFe(C2O4)x as a heterogeneous Fenton catalyst for removal of methyl orange.

This paper studies a heterogeneous Fenton catalyst NiFe(C2O4)x, which showed better catalytic activity than Ni(C2O4)x and better re-usability than Fe(C2O4)x. The methyl orange removal efficiency was 98% in heterogeneous Fenton system using NiFe(C2O4)x. The prepared NiFe(C2O4)x had a laminated shape and the size was in the range of 2-4 µm, and Ni was doped into catalyst's structure successfully. The NiFe(C2O4)x had a synergistic effect of catalyst of 24.7 for methyl orange removal, and the dope of Ni significantly reduced the leaching of Fe by 77%. The reaction factors and kinetics were investigated. Under the optimal conditions, 0.4 g/L of catalyst dose and 10 mmol/L of hydrogen peroxide concentration, 98% of methyl orange was removed within 20 min. Analysis showed that hydroxyl radicals and superoxide radicals participated in the reaction. With NiFe(C2O4)x catalyst, the suitable pH range for heterogeneous Fenton system was wide from 3 to 10. The catalyst showed good efficiency after five times re-use. NiFe(C2O4)x provided great potential in treatment of refractory wastewater with excellent property.

Degradation of aniline by heterogeneous Fenton's reaction using a Ni-Fe oxalate complex catalyst.

A Ni-Fe oxalate complex catalyst was synthesized and characterized by means of Brunauer-Emmet-Teller (BET) method, scanning electron microscope (SEM) and X-ray photo-electron spectroscopy (XPS). The catalyst showed good catalytic activity for aniline degradation by heterogeneous Fenton's reaction, in which the synergetic index was 9.3. The effects of reaction temperature, catalyst dosage, hydrogen peroxide concentration and initial pH were investigated. Under the optimum conditions (T = 293 K, catalyst dosage = 0.2 g/L, H2O2 concentration = 4 mmol/L and initial pH = 5.4), 100% aniline could be removed within 35 min, and approximately 88% deamination efficiency was achieved in 60 min. The aniline degradation process followed the pseudo-first-order kinetic (k = 0.177 min(-1)) with activation energy (Ea) of 49.4 kJ mol(-1). Aniline could be removed in a broad initial pH (3-8) due to the excellent pH-tolerance property of the catalyst. The detected ammonium ion indicated that deamination occurred during aniline degradation. It was proposed that deamination synchronized with aniline removal, and aniline was attacked by free radicals to generate benzoquinonimine and phenol. This system is promising for the removal of aniline from water.

Preparation of FeCeOx by ultrasonic impregnation method for heterogeneous Fenton degradation of diclofenac.

FeCeOx has been successfully synthesized by ultrasonic impregnation method and applied in diclofenac removal in heterogeneous Fenton process. The effects of ultrasonic density, impregnation time, mole ratio of Fe and Ce and calcination temperature were investigated. Nitrogen adsorption/desorption, SEM, XRD, HRTEM, Raman and XPS analyses were characterized. Stability and reusability of FeCeOx were evaluated. The results indicated that 83% degradation efficiency of diclofenac was achieved by FeCeOx under the optimum preparation conditions. Fe ions were distributed uniformly in crystal structure and the solid solution structure of FeCeOx with a lattice constriction was formed. Exposed crystalline plane (200) with a relatively high surface energy may be the main reason to provide high catalytic activity of FeCeOx. Oxygen vacancies took part in catalytic process and a portion of them were oxidized after reaction. FeCeOx showed an excellent chemical stability and reusability in heterogeneous Fenton process.