Biosensors — Open Access Journal

Bacteria in the genus Brucella are the cause of brucellosis in humans and many domestic and wild animals. A rapid and culture-free detection assay to detect Brucella in clinical samples would be highly valuable. Nanomaterial optical fiber biosensors (NOFS) are capable of recognizing DNA hybridization events or other analyte interactions with high specificity and sensitivity. Therefore, a NOFS assay was developed to detect Brucella DNA from cultures and in tissue samples from infected mice. An ionic self-assembled multilayer (ISAM) film was coupled to a long-period grating optical fiber, and a nucleotide probe complementary to the Brucella IS711 region and modified with biotin was bound to the ISAM by covalent conjugation. When the ISAM/probe duplex was exposed to lysate containing ≥100 killed cells of Brucella, or liver or spleen tissue extracts from Brucella-infected mice, substantial attenuation of light transmission occurred, whereas exposure of the complexed fiber to non-Brucella gram-negative bacteria or control tissue samples resulted in negligible attenuation of light transmission. Oligonucleotide probes specific for B. abortus, B. melitensis, and B. suis could also be used to detect and differentiate these three nomenspecies. In summary, the NOFS biosensor assay detected three nomenspecies of Brucella without the use of polymerase chain reaction within 30 min and could specifically detect low numbers of this bacterium in clinical samples. Full article

A biosensitive element in the form of bacterial Photobacterium phosphoreum cells immobilized in poly(vinyl alcohol) cryogel was tested for the determination of different mycotoxins under discrete and flow-through analysis conditions. The immobilized bioluminescent cells made it possible to quantify the presence of Ochratoxin A, Sterigmatocystin, Zearalenone, and Deoxynivalenon in aqueous media in a wide range of their concentrations (0.017–56 mg/L, 0.010–33 mg/L, 0.009–14 mg/L, and 0.026–177 mg/L, respectively) via measuring the quenching of cell luminescence. The flow conditions allowed the analysis sensitivity to be improved by an order of magnitude in terms of detected concentrations. Using the immobilized luminescent bacterial cells, we have shown the possibility of evaluating the efficiency of the mycotoxins’ hydrolysis under the action of enzymes. In this way, a 94 ± 4.5% efficiency of Zearalenone hydrolysis with hexahistidine-containing organophosphorus hydrolase for 1h-long treatment of the mycotoxin solution (100 mg/L) was shown. Full article

In the developing world, the identification of clean, potable water continues to pose a pervasive challenge, and waterborne diseases due to fecal contamination of water supplies significantly threaten public health. The ability to efficiently monitor local water supplies is key to water safety, yet no low-cost, reliable method exists to detect contamination quickly. We developed an in vitro assay utilizing an odorant-binding protein (OBP), AgamOBP1, from the mosquito, Anopheles gambiae, to test for the presence of a characteristic metabolite, indole, from harmful coliform bacteria. We demonstrated that recombinantly expressed AgamOBP1 binds indole with high sensitivity. Our proof-of-concept assay is fluorescence-based and demonstrates the usefulness of insect OBPs as detector elements in novel biosensors that rapidly detect the presence of bacterial metabolic markers, and thus of coliform bacteria. We further demonstrated that rAgamOBP1 is suitable for use in portable, inexpensive “dipstick” biosensors that improve upon lateral flow technology since insect OBPs are robust, easily obtainable via recombinant expression, and resist detector “fouling.” Moreover, due to their wide diversity and ligand selectivity, insect chemosensory proteins have other biosensor applications for various analytes. The techniques presented here therefore represent platform technologies applicable to various future devices. Full article

Antibiotics are an important class of drugs destined for treatment of bacterial diseases. Misuses and overuses of antibiotics observed over the last decade have led to global problems of bacterial resistance against antibiotics (ABR). One of the crucial actions taken towards limiting the spread of antibiotics and controlling this dangerous phenomenon is the sensitive and accurate determination of antibiotics residues in body fluids, food products, and animals, as well as monitoring their presence in the environment. Immunosensors, a group of biosensors, can be considered an attractive tool because of their simplicity, rapid action, low-cost analysis, and especially, the unique selectivity arising from harnessing the antigen–antibody interaction that is the basis of immunosensor functioning. Herein, we present the recent achievements in the field of electrochemical immunosensors designed to determination of antibiotics. Full article

In a simple and instant procedure for detecting fish freshness, a hydrogel and hydrophilic pectin matrix membrane was used successfully as an optical pH sensor by immobilizing the chromoionophore ETH 5294 (CI), which is very selective and sensitive for the membrane. The Pe/CI optical pH sensor exhibited excellent linearity between pH 5 and pH 9, with a sensor response time of 5 min and reproducibility of 1.49% relative standard deviation (RSD). The sensor showed response stability for 15 days and a response reduction of 8.6%. The sensor’s capability was demonstrated by the detection of fish freshness for 17 days at 4 °C. Full article

This study investigated the difference in the gait of patients with Parkinson’s disease (PD), age-matched controls and young controls during three walking patterns. Experiments were conducted with 24 PD, 24 age-matched controls and 24 young controls, and four gait intervals were measured using inertial measurement units (IMU). Group differences between the mean and variance of the gait parameters (stride interval, stance interval, swing interval and double support interval) for the three groups were calculated and statistical significance was tested. The results showed that the variance in each of the four gait parameters of PD patients was significantly higher compared with the controls, irrespective of the three walking patterns. This study showed that the variance of any of the gait interval parameters obtained using IMU during any of the walking patterns could be used to differentiate between the gait of PD and control people. Full article

With the rapid increase in the development of miniaturized sensors and embedded devices for vital signs monitoring, personal physiological signal monitoring devices are becoming popular. However, physiological monitoring devices which are worn on the body normally affect the daily activities of people. This problem can be avoided by using a non-contact measuring device like the Doppler radar system, which is more convenient, is private compared to video monitoring, infrared monitoring and other non-contact methods. Additionally real-time physiological monitoring with the Doppler radar system can also obtain signal changes caused by motion changes. As a result, the Doppler radar system not only obtains the information of respiratory and cardiac signals, but also obtains information about body movement. The relevant RF technology could eliminate some interference from body motion with a small amplitude. However, the motion recognition method can also be used to classify related body motion signals. In this paper, a vital sign and body movement monitoring system worked at 2.4 GHz was proposed. It can measure various physiological signs of the human body in a non-contact manner. The accuracy of the non-contact physiological signal monitoring system was analyzed. First, the working distance of the system was tested. Then, the algorithm of mining collective motion signal was classified, and the accuracy was 88%, which could be further improved in the system. In addition, the mean absolute error values of heart rate and respiratory rate were 0.8 beats/min and 3.5 beats/min, respectively, and the reliability of the system was verified by comparing the respiratory waveforms with the contact equipment at different distances. Full article

Surface-enhanced Raman scattering (SERS) has become a powerful tool in chemical, material and life sciences, owing to its intrinsic features (i.e., fingerprint recognition capabilities and high sensitivity) and to the technological advancements that have lowered the cost of the instruments and improved their sensitivity and user-friendliness. We provide an overview of the most significant aspects of SERS. First, the phenomena at the basis of the SERS amplification are described. Then, the measurement of the enhancement and the key factors that determine it (the materials, the hot spots, and the analyte-surface distance) are discussed. A section is dedicated to the analysis of the relevant factors for the choice of the excitation wavelength in a SERS experiment. Several types of substrates and fabrication methods are illustrated, along with some examples of the coupling of SERS with separation and capturing techniques. Finally, a representative selection of applications in the biomedical field, with direct and indirect protocols, is provided. We intentionally avoided using a highly technical language and, whenever possible, intuitive explanations of the involved phenomena are provided, in order to make this review suitable to scientists with different degrees of specialization in this field. Full article

Electric cell-substrate impedance sensing (ECIS) is an impedance-based method for monitoring changes in cell behaviour in real-time. In this paper, we highlight the importance of ECIS in measuring the kinetics of human melanoma cell invasion across human brain endothelium. ECIS data can be mathematically modelled to assess which component of the endothelial paracellular and basolateral barriers is being affected and when. Our results reveal that a range of human melanoma cells can mediate disruption of human brain endothelium, primarily involving the paracellular route, as demonstrated by ECIS. The sensitivity of ECIS also reveals that the paracellular barrier weakens within 30–60 min of the melanoma cells being added to the apical face of the endothelial cells. Imaging reveals pronounced localisation of the melanoma cells at the paracellular junctions consistent with paracellular migration. Time-lapse imaging further reveals junctional opening and disruption of the endothelial monolayer by the invasive melanoma cells all within several hours. We suggest that the ability of ECIS to resolve changes to barrier integrity in real time, and to determine the route of migration, provides a powerful tool for future studies investigating the key molecules involved in the invasive process of cancer cells. Full article

Early diagnosis of inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), remains a clinical challenge with current tests being invasive and costly. The analysis of volatile organic compounds (VOCs) in exhaled breath and biomarkers in stool (faecal calprotectin (FCP)) show increasing potential as non-invasive diagnostic tools. The aim of this pilot study is to evaluate the efficacy of breath analysis and determine if FCP can be used as an additional non-invasive parameter to supplement breath results, for the diagnosis of IBD. Thirty-nine subjects were recruited (14 CD, 16 UC, 9 controls). Breath samples were analysed using an in-house built electronic nose (Wolf eNose) and commercial gas chromatograph–ion mobility spectrometer (G.A.S. BreathSpec GC-IMS). Both technologies could consistently separate IBD and controls [AUC ± 95%, sensitivity, specificity], eNose: [0.81, 0.67, 0.89]; GC-IMS: [0.93, 0.87, 0.89]. Furthermore, we could separate CD from UC, eNose: [0.88, 0.71, 0.88]; GC-IMS: [0.71, 0.86, 0.62]. Including FCP did not improve distinction between CD vs. UC; eNose: [0.74, 1.00, 0.56], but rather, improved separation of CD vs. controls and UC vs. controls; eNose: [0.77, 0.55, 1.00] and [0.72, 0.89, 0.67] without FCP, [0.81, 0.73, 0.78] and [0.90, 1.00, 0.78] with FCP, respectively. These results confirm the utility of breath analysis to distinguish between IBD-related diagnostic groups. FCP does not add significant diagnostic value to breath analysis within this study. Full article

Development of technologies for rapid screening of DNA secondary structure thermal stability and the effects on stability for binding of small molecule drugs is important to the drug discovery process. In this report, we describe the capabilities of an electrochemical, microdevice-based approach for determining the melting temperatures (T_m) of electrode-bound duplex DNA structures. We also highlight new features of the technology that are compatible with array development and adaptation for high-throughput screening. As a foundational study to exhibit device performance and capabilities, melting-curve analyses were performed on 12-mer DNA duplexes in the presence/absence of two binding ligands: diminazene aceturate (DMZ) and proflavine. By measuring electrochemical current as a function of temperature, our measurement platform has the ability to determine the effect of binding ligands on T_m values with high signal-to-noise ratios and good reproducibility. We also demonstrate that heating our three-electrode cell with either an embedded microheater or a thermoelectric module produces similar results. The ΔT_m values we report show the stabilizing ability of DMZ and proflavine when bound to duplex DNA structures. These initial proof-of-concept studies highlight the operating characteristics of the microdevice platform and the potential for future application toward other immobilized samples. Full article

In this study, stable gold nanoparticles (AuNPs) are fabricated for the first time on commercial ultrafine glass coverslips coated with gold thin layers (2 nm, 4 nm, 6 nm, and 8 nm) at 25 °C and annealed at high temperatures (350 °C, 450 °C, and 550 °C) on a hot plate for different periods of time. Such gold nanostructured coverslips were systematically tested via surface enhanced Raman spectroscopy (SERS) to identify their spectral performances in the presence of different concentrations of a model molecule, namely 1,2-bis-(4-pyridyl)-ethene (BPE). By using these SERS platforms, it is possible to detect BPE traces (10⁻¹² M) in aqueous solutions in 120 s. The stability of SERS spectra over five weeks of thiol-DNA probe (2 µL) deposited on gold nano-structured coverslip is also reported. Full article

Conjugation chemistry does not always provide adequate spatial orientation of hapten in immunogens for the best presentation of generic or individual epitopes. In the present study, the influence of unique and multiple orientations of immunizing hapten on the immune response repertoire was compared to select generic recognition system. The glycopeptides, teicoplanin (TPL) and ristomycin (RSM), were conjugated to BSA to produce immunogens with unique and multiple orientations of haptens. Polyclonal antibodies generated against TPL conjugated through a single site were of uniform specificity and demonstrated selective TPL recognition, regardless of the coating conjugates design. The sensitivity (IC₅₀) of 4 enzyme-linked immunosorbent assays (ELISAs) for TPL varied little within the 3.5–7.4 ng/mL, with a dynamic range of 0.2–100 ng/mL. RSM was coupled to BSA through several glycoside sites that evoked a wider repertoire of response. This first described anti-RSM antibody was selective for RSM in homologous hapten-coated ELISAs with IC₅₀ values in the range 4.2–35 ng/mL. Among the heterologous antigens, periodate-oxidized TPL conjugated to gelatine was selected as the best binder of generic anti-RSM fraction. The developed ELISA showed group recognition of glycopeptides RSM, TPL, eremomycin, and vancomycin with cross-reactivity of 37–100% and a 10–10,000 ng/mL dynamic range. Thus, multiple presentations of immunizing hapten help expand the repertoire of immune responses and opportunities for the selection of the required fine-specificity agent. Full article

We have designed and validated a set of robust and non-toxic protocols for directly evaluating the properties of engineered neural tissue. These protocols characterize the mechanical properties of engineered neural tissues and measure their electrophysical activity. The protocols obtain elastic moduli of very soft fibrin hydrogel scaffolds and voltage readings from motor neuron cultures. Neurons require soft substrates to differentiate and mature, however measuring the elastic moduli of soft substrates remains difficult to accurately measure using standard protocols such as atomic force microscopy or shear rheology. Here we validate a direct method for acquiring elastic modulus of fibrin using a modified Hertz model for thin films. In this method, spherical indenters are positioned on top of the fibrin samples, generating an indentation depth that is then correlated with elastic modulus. Neurons function by transmitting electrical signals to one another and being able to assess the development of electrical signaling serves is an important verification step when engineering neural tissues. We then validated a protocol wherein the electrical activity of motor neural cultures is measured directly by a voltage sensitive dye and a microplate reader without causing damage to the cells. These protocols provide a non-destructive method for characterizing the mechanical and electrical properties of living spinal cord tissues using novel biosensing methods. Full article

A highly sensitive electrochemical biosensor with a signal amplification platform of electrodeposited gold nanoparticle (AuNP) has been developed and characterized. The sizes of the synthesized AuNP were found to be critical for the performance of biosensor in which the sizes were dependent on HAuCl₄ and acid concentrations; as well as on scan cycles and scan rates in the gold electro-reduction step. Systematic investigations of the adsorption of proteins with different sizes from aqueous electrolyte solution onto the electrodeposited AuNP surface were performed with a potentiometric method and calibrated by design of experiment (DOE). The resulting amperometric glucose biosensors was demonstrated to have a low detection limit (>50 μM) and a wide linear range after optimization with AuNP electrodeposition. Full article

With brain tumour incidence increasing, there is an urgent need for better diagnostic tools. Intraoperatively, brain tumours are diagnosed using a smear preparation reported by a neuropathologist. These have many limitations, including the time taken for the specimen to reach the pathology department and for results to be communicated to the surgeon. There is also a need to assist with resection rates and identifying infiltrative tumour edges intraoperatively to improve clearance. We present a novel study using a handheld Raman probe in conjunction with gold nanoparticles, to detect primary and metastatic brain tumours from fresh brain tissue sent for intraoperative smear diagnosis. Fresh brain tissue samples sent for intraoperative smear diagnosis were tested using the handheld Raman probe after application of gold nanoparticles. Derived Raman spectra were inputted into forward feature extraction algorithms to build a predictive model for sensitivity and specificity of outcome. These results demonstrate an ability to detect primary from metastatic tumours (especially for normal and low grade lesions), in which accuracy, sensitivity and specificity were respectively equal to 98.6%, 94.4% and 99.5% for normal brain tissue; 96.1%, 92.2% and 97.0% for low grade glial tumours; 90.3%, 89.7% and 90.6% for high grade glial tumours; 94.8%, 63.9% and 97.1% for meningiomas; 95.4%, 79.2% and 98.8% for metastases; and 99.6%, 88.9% and 100% for lymphoma, based on smear samples (κ = 0.87). Similar results were observed when compared to the final formalin-fixed paraffin embedded tissue diagnosis (κ = 0.85). Overall, our results have demonstrated the ability of Raman spectroscopy to match results provided by intraoperative smear diagnosis and raise the possibility of use intraoperatively to aid surgeons by providing faster diagnosis. Moving this technology into theatre will allow it to develop further and thus reach its potential in the clinical arena. Full article

This paper demonstrated a new design structure for minimizing the power consumption of a pulse meter. Monolithic devices composed of a red (625 nm) organic light-emitting diode (OLED) and an organic photodiode (OPD) were fabricated on the same substrate. Two organic devices were designed differently. One had a circle-shaped OLED in the center of the device and was surrounded by the OPD, while the other had the opposite structure. The external quantum efficiency (EQE) of the OLED and the OPD were 7% and 37%, respectively. We evaluated and compared the signal-to-noise ratio (SNR) of the photoplethysmogram (PPG) signal on different parts of the body and successfully acquired clear PPG signals at those positions, where the best signal was obtained from the fingertip at a SNR of about 62 dB. The proposed organic pulse meter sensor was operated successfully with a power consumption of 0.1 mW. Eventually, the proposed organic biosensor reduced the power consumption and improved the capability of the pulse meter for long-term use. Full article

Innovative methods to achieve the user-friendly, quick, and highly sensitive detection of nanomaterials are urgently needed. Nanomaterials have increased importance in commercial products, and there are concerns about the potential risk that they entail for the environment. In addition, detection of nanomaterials can be a highly valuable tool in many applications, such as biosensing. Electrochemical methods using disposable, low-cost, printed electrodes provide excellent analytical performance for the detection of a wide set of nanomaterials. In this review, the foundations and latest advances of several electrochemical strategies for the detection of nanoparticles using cost-effective printed devices are introduced. These strategies will equip the experimentalist with an extensive toolbox for the detection of nanoparticles of different chemical nature and possible applications ranging from quality control to environmental analysis and biosensing. Full article

Continuous progress in the domain of nano and material science has led to modulation of the properties of nanomaterials in a controlled and desired fashion. In this sense, nanomaterials, including carbon-based materials, metals and metal oxides, and composite/hybrid materials have attracted extensive interest with regard to the construction of electrochemical biosensors. The modification of a working electrode with a combination of two or three nanomaterials in the form of nano-composite/nano-hybrids has revealed good results with very good reproducibility, stability, and improved sensitivity. This review paper is focused on discussing the possible constructs of nano-hybrids and their subsequent use in the construction of electrochemical glucose biosensors. Full article