Low-cost Biosensor Research Articles

Protein-Templated Click Ligation Reaction Triggered by Protein-Split Aptamer Interactions.

DNA-templated reactions have found wide applications in sensing and drug discovery. However, this strategy has been limited to the use of nucleic acids as templating elements to direct the proximity effect. Herein, we describe a versatile protein-templated split aptamer click ligation reaction (PT-SpA-CLR) in which the protein template-induced covalent proximity ligation of split aptamer elements enables translating protein/aptamer binding events into the output of ligated DNA products. A ligation yield of >80% is observed for three model protein templates, including VEGF165, PDGF-BB, and SARS-CoV-2 S1. The ligation reaction compensates for the weakness of reduced binding affinity resulting from splitting the aptamer, as evidenced by an approximately 2-fold lower dissociation constant than the non-ligated system. This newly developed PT-SpA-CLR strategy is further integrated with colorimetric or fluorescent reporting mechanisms to achieve easy-to-use and low-cost biosensors utilizing ligation to produce a fully active G-quadruplex or an RNA-cleaving DNAzyme to report protein binding. Both assays can achieve specific detection of an intended protein target with a limit of detection at the picomolar level even when challenged in biological samples. The combined PT-SpA-CLR and versatile sensing strategies offer attractive universal platforms for efficient detection of protein biomarkers.

Enhancing food safety: A low-cost biosensor for Bacillus licheniformis detection in food products

To enhance food safety, the need for swift and precise detection of B. licheniformis, a bacterium prevalent in various environments, including soil and food products, is paramount. This study presents an innovative and cost-effective bioassay designed to specifically identify the foodborne pathogen, B. licheniformis, utilizing a colorimetric signal approach. The biosensor, featuring a 3D-printed architecture, incorporates a casein-based liquid-proof gelatine film, selectively liquefying in response to the caseinolytic/proteolytic activity of external enzymes from the pathogen. As the sample liquefies, it progresses through a color layer, causing the migration of dye to an absorbent layer, resulting in a distinct positive signal. This bioassay exhibits exceptional sensitivity, detecting concentrations as low as 1 CFU/mL within a 9.3-h assay duration. Notably, this cost-efficient bioassay outperforms conventional methods in terms of efficacy and cost-effectiveness, offering a straightforward solution for promptly detecting B. licheniformis in food samples.

Low-Cost Biosensor Technologies for Rapid Detection of COVID-19 and Future Pandemics.

Many systems have been designed for the detection of SARS-CoV-2, which is the virus that causes COVID-19. SARS-CoV-2 is readily transmitted, resulting in the rapid spread of disease in human populations. Frequent testing at the point of care (POC) is a key aspect for controlling outbreaks caused by SARS-CoV-2 and other emerging pathogens, as the early identification of infected individuals can then be followed by appropriate measures of isolation or treatment, maximizing the chances of recovery and preventing infectious spread. Diagnostic tools used for high-frequency testing should be inexpensive, provide a rapid diagnostic response without sophisticated equipment, and be amenable to manufacturing on a large scale. The application of these devices should enable large-scale data collection, help control viral transmission, and prevent disease propagation. Here we review functional nanomaterial-based optical and electrochemical biosensors for accessible POC testing for COVID-19. These biosensors incorporate nanomaterials coupled with paper-based analytical devices and other inexpensive substrates, traditional lateral flow technology (antigen and antibody immunoassays), and innovative biosensing methods. We critically discuss the advantages and disadvantages of nanobiosensor-based approaches compared to widely used technologies such as PCR, ELISA, and LAMP. Moreover, we delineate the main technological, (bio)chemical, translational, and regulatory challenges associated with developing functional and reliable biosensors, which have prevented their translation into the clinic. Finally, we highlight how nanobiosensors, given their unique advantages over existing diagnostic tests, may help in future pandemics.

Portable biosensor combining CRISPR/Cas12a and loop-mediated isothermal amplification for antibiotic resistance gene ermB in wastewater

Wastewater is among the main sources of antibiotic resistance genes (ARGs) in the environment, but effective methods to quickly assess ARGs on-site in wastewater are lacking. Here, using the typical ARG ermB as the target, we report a portable biosensor combining CRISPR/Cas12a and loop-mediated isothermal amplification (LAMP) for the detection of ARGs. Six primers of LAMP and the crRNA of CRISPR/Cas12a were first designed to be preamplification with LAMP and lead Cas12a to recognize the ermB via base pairing. Due to the trans-cleavage activity of CRISPR/Cas12a after amplicon recognition, ssDNA probes modified with reporter molecules were used to implement a visual assay with lateral flow test strips and fluorescence. After a simple nucleic acid extraction with magnetic beads, the constructed biosensor possesses excellent sensitivity and selectivity as low as 2.75 × 103 copies/μL using fluorescence and later flow strips in wastewater. We further evaluated the community-wide prevalence of ermB in wastewater influent and found high mass loads of ermB during different months. This user-friendly and low-cost biosensor is applicable for rapid on-site ARG detection, providing a potential point-of-use method for rapid assessments of ARG abundance in wastewater from large city areas with many wastewater treatment plants and in resource-limited rural areas.

Analysis of Plasmonic Sensors Performance Realized by Exploiting Different UV-Cured Optical Adhesives Combined with Plastic Optical Fibers.

Polymer-based surface plasmon resonance (SPR) sensors can be used to realize simple, small-size, disposable, and low-cost biosensors for application in several fields, e.g., healthcare. The performance of SPR sensors based on optical waveguides can be changed by tuning several parameters, such as the dimensions and the shape of the waveguides, the refractive index of the core, and the metal nanofilms used to excite the SPR phenomenon. In this work, in order to develop, experimentally test, and compare several polymer-based plasmonic sensors, realized by using waveguides with different core refractive indices, optical adhesives and 3D printed blocks with a trench inside have been used. In particular, the sensors are realized by filling the blocks' trenches (with two plastic optical fibers located at the end of these) with different UV-cured optical adhesives and then covering them with the same bilayer to excite the SPR phenomenon. The developed SPR sensors have been characterized by numerical and experimental results. Finally, in order to propose photonic solutions for healthcare, a comparative analysis has been reported to choose the best sensor configuration useful for developing low-cost biosensors.

Microelectromechanical system‐based biosensor for label‐free detection of human cytomegalovirus

The human cytomegalovirus (HCMV) is an asymptomatic common virus that is typically harmless, but in some cases, it can be life threatening. Thus, there is an urgent need to develop novel diagnostic methods and strengthen the efforts to combat this virus. A microcantilever-based biosensor functionalised with the UL83-antibody of HCMV (UL83-HCMV antibody) has been developed to detect the UL83-antigen of HCMV (UL83-HCMV antigen) at different concentrations ranging from 0.3 to 300ng/ml. The response of the biosensor to the presence of UL83-HCMV antigen was measured through the shift in resonance frequency before and after antigen-antibody binding. The system shows a low detection limit of 84pg/ml, which is comparable to traditional sensors, and a detection time of less than 15min was achieved. The selectivity of the sensor was demonstrated using three different proteins with and without the UL83-HCMV antigen. The biosensor shows high selectivity for the UL83-HCMV antigen. Mass loading by the UL83-HCMV antigen was roughly estimated with a sensitivity of ∼30fg/Hz. This technique is crucial for the fabrication of portable and low-cost biosensors that can be used in real-time monitoring and enables early medical diagnosis.

Gold Nanoparticles-MWCNT Based Aptasensor for Early Diagnosis of Prostate Cancer.

Prostate cancer is one of the most frequently diagnosed male malignancies and can be detected by prostate-specific antigen (PSA) as a biomarker. To detect PSA, several studies have proposed using antibodies, which are not economical and require a long reaction time. In this study, we propose to use self-assembled thiolated single-strand DNA on electrodes functionalized by multi-walled carbon nanotubes (MWCNT) modified with gold nanoparticles (AuNPs) to realize a low-cost label-free electrochemical biosensor. In this regard, the PSA aptamer was immobilized via electrostatic adsorption on the surface of a screen-printed MWCNT/AuNPs electrode. The immobilization process was enhanced due to the presence of Au nanoparticles on the surface of the electrode. Surface characterization of the electrode at different stages of modification was performed by electrochemical impedance spectroscopy (EIS), atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) and contact angle for surface tension properties. The results showed an increase in surface roughness due to the absorbance of the aptamer on the electrode surfaces. The developed sensor has an extended linear range of 1-100 ng/mL, and a very low limit of detection down to 1 pg/mL. In addition, the reaction has a binding time of only five minutes on the developed electrodes. Investigations of the biosensor selectivity against several substances revealed an efficient selectivity for PSA detection. With this approach, low-cost biosensors with high sensitivity can be realized which have a wide linearity range and a low limit of detection, which are necessary for the early detection of prostate cancer.

A Noninvasive Sweat Glucose Biosensor Based on Glucose Oxidase/Multiwalled Carbon Nanotubes/Ferrocene-Polyaniline Film/Cu Electrodes.

Diabetes remains a great threat to human beings' health and its world prevalence is projected to reach 9.9% by 2045. At present, the detection methods used are often invasive, cumbersome and time-consuming, thus increasing the burden on patients. In this paper, we propose a novel noninvasive and low-cost biosensor capable of detecting glucose in human sweat using enzyme-based electrodes for point-of-care uses. Specifically, an electrochemical method is applied for detection and the electrodes are covered with multilayered films including ferrocene-polyaniline (F-P), multi-walled carbon nanotubes (MWCNTs) and glucose oxidase (GOx) on Cu substrates (GOx/MWCNTs/F-P/Cu). The coated layers enhance the immobilization of GOx, increase the conductivity of the anode and improve the electrochemical properties of the electrode. Compared with the Cu electrode and the F-P/Cu electrode, a maximum peak current is obtained when the MWCNTs/F-P/Cu electrode is applied. We also study its current response by cyclic voltammetry (CV) at different concentrations (0-2.0 mM) of glucose solution. The best current response is obtained at 0.25 V using chronoamperometry. The effective working lifetime of an electrode is up to 8 days. Finally, to demonstrate the capability of the electrode, a portable, miniaturized and integrated detection device based on the GOx/MWCNTs/F-P/Cu electrode is developed. The results exhibit a short response time of 5 s and a correlation coefficient R2 of 0.9847 between the response current of sweat with blood glucose concentration. The LOD is of 0.081 mM and the reproducibility achieved in terms of RSD is 3.55%. The sweat glucose sensor is noninvasive and point-of-care, which shows great development potential in the health examination and monitoring field.

(Invited, Digital Presentation) Super-Nernstian Isfet Combining Two-Dimensional WSe2/MoS2 Heterostructure with Negative Capacitance

Ion-sensitive field-effect transistors (ISFETs) are quite popular as compact, low-cost biosensors with fast response time and label-free detection1. They can be used as pH sensors or functionalized for complex biomolecule detection. The voltage sensitivity (Sv) in classical ISFETs is fundamentally limited to 59 mV/pH (Nernst limit). Surpassing the Nernst limit requires complex device architectures or novel transport phenomena. Sensitivity beyond the Nernst limit can be achieved using specific device architectures such as dual gate ISFETs2, negative capacitance ISFETs (NC-ISFET)3, tunnel ISFETs4, etc. Compatible architectures can be combined for further enhancements in sensitivity.First, we experimentally demonstrate a super-Nernstian hetero-ISFET that uses 2-D WSe2/MoS2 heterostructure in a double-gated configuration5. The schematic of the device structure is shown in Fig. 1(a) along with its dimensions. The fluid gate to the pH solution is biased at VFG = 0 V and the voltage sensitivity (SV) is extracted by applying bias to the back-gate (VBG). Fig. 1(b) shows the variation of drain current for change in VBG at different pH. The voltage sensitivity is also included in the same graph. The device uses charge screening due to the interface traps and inversion charges at the hetero-interface to modulate the back-gate transconductance (gmb), thereby allowing super sensitivity.Further enhancement in sensitivity is explored using technology computer-aided (TCAD) device simulator tool (Silvaco ATLAS) by integrating with different device architectures. First we model the baseline hetero-ISFET. The 2-D materials were modeled using their material parameters and 3-D equivalents of their density of states. Amorphous hafnium oxide (HfO2) was used as the dielectric. The mobile ions in the electrolyte were modeled as charge carriers in an intrinsic semiconductor, with its effective density of states varying as a function of pH. The simulation model was calibrated with the experimental device (at pH = 7), as shown in Fig. 2(a). The transfer characteristics of the back-gate at different pH and fixed VFG (= 0 V) for the simulated device is shown in Fig. 2(b). We note that the sensitivity from simulations is lower than the experimental device. This is likely due to non-ideal and 2-D material specific factors which are not accounted in simulations. Nevertheless, the simulated device also shows super-Nernstian sensitivity (Fig. 2(b), right axis), validating the model. Hence, the calibrated TCAD model is used as the baseline for further studies.Next, an NC-FE layer (aluminum-doped HfO2) was added to the top fluid-gate stack6. We have used a ferroelectric-metal-insulator-semiconductor (FMIS) stack for the proposed NC-hetero-ISFET. Fig. 3(a) shows the new top-gate stack with the FMI layer, which replaces the top-gate stack in the earlier schematic. The fluid-gate charge (QFG), and drain current (ID) as a function of VFG (VBG = 0 V), were obtained from the TCAD simulations. The 1-D Landau–Khalatnikov (L-K) equations were used to model the voltage across the FE layer (VFE = 2αQFG+4βQ3 FG = V' FG - Vint; where V' FG is the newly computed fluid-gate bias and Vint is the internal node voltage)7. The calculated Vint (for fixed V' FG) is coupled back into the ATLAS simulator to extract voltage sensitivity (SV) by sweeping VBG at different pH values.The fluid-gate transfer curve of the proposed NC-hetero-ISFET, in Fig. 3(b), clearly shows a steeper sub-threshold slope and higher ON current than the baseline device. The corresponding FE layer parameters are shown in Table 1. These improved fluid-gate characteristics contribute to an increased voltage-sensitivity (SV) when VBG is applied. The transfer characteristic (ID v/s VBG, at fixed V' FG) of the NC-hetero-ISFET at different pH values is shown in Fig. 3(c), along with the voltage sensitivity. Further, in Fig. 3(d), we compare the peak SV obtained at different V' FG. There is an improvement in voltage sensitivity (as much as ~ 100 mV/pH) over the baseline device when NC is introduced. The results pave the way for highly sensitive super-Nernstian ISFETs by combining 2-D heterostructure with NC effect.

Development of Selective Cortisol Biosensors in the Control of Fatigue and Related Diseases

The study of fatigue becomes complex due to several subjective factors related to its causality, making the relationship between the level of fatigue and the concentration of cortisol in an individual characteristic. However, cortisol is identified as the main element released in response to fatigue, and as such, it can be used as a biomarker for its determination and other associated diseases. Inadequate amounts of cortisol are related to non-specific diseases such as diabetes and osteoporosis. The body's cortisol levels fluctuate throughout the day, with the greatest levels in the morning and the lowest in the evening. To correlate these various fluctuations to the degree of fatigue, is required a real-time determination of cortisol levels with an effective sensory device, which overcomes the demand of costly and time-consuming laboratory analysis. Cortisol, as expressed in sweat, has a wide dynamic concentration range that can be used to monitor fatigue during different activities, being useful in preventing diseases and accidents. Currently, cortisol evaluation may be critical as one of the prospective biomarkers for Covid-19-induced significant inflammatory consequences. Some studies show possible severity between the production of pro-inflammatory cytokines related to viral infection by the new Sars-Cov-2 in people with above-average serum cortisol levels. In this regard, the establishment of a simple and low-cost biosensor with rapid diagnosis and in loco application would be beneficial due to the analytical center's independence and the simplicity with patients positive for the Sars-Cov-2 virus or others may be followed and treated. For the cortisol biosensor development, electrochemically exfoliated graphene (EEG) was used as a current collector nanomaterial in combination with chitosan (CS), a biomaterial derived from discarded crustacean shells. The carbon DropSens were modified with nanocomposite containing CS and EEG prepared using an ultrasonication. 21µL of CS_EEG dispersion by drop casting and exposed for 4 hours by white light, in order to dry the nanocomposite and recover all the working electrode. The material not bounded to electrode was then removed after 3 washes of 50µL of Milli-Q water. 10 µL of glutaraldehyde 1 % solution (GA) was dripped on the electrodes and allowed to react for 15 minutes to guarantee greater interaction of the nanocomposite with the antibody. The wet surface of electrodes was then evaluated for the incorporation of monoclonal antibody solution by IgG type specific for cortisol at varying concentrations. A total of 7.5 µL of diluted IgG solution (3.33 ng mL-1) and pure IgG solution were added to the biosensor immune complex (100 ng mL-1) and allowed to act for 30 minutes. Sensory measurements are performed in standard cortisol dilutions in PBS 7.4 in the following concentrations: 1 ng mL-1, 10ng mL -1, 100ng mL-1, 200ng mL-1, 500ng mL-1 and 1 µg. Then, the electrodes with and without antibody were characterized, via electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), FEG microscopy, Goniometry and Zeta Potential. Goniometry assays showed an increase of hydrophilic profile of the nanocomposites after antibody modification conferring a suit surface for biological elements deposition, Fig 1. FTIR and FEG Microscopy provided the interaction of antibody-nanocomposite and antibody- antigen complex, Fig 2 and 3. Infrared analysis showed bands assigned to methyl groups disappeared after antibody adhesion and this condition may relates to the recovery of heavy chains of IgG on the nanocomposite. The increase in capacitance value after each sensory electrochemical measurement provided a cortisol curve calibration, Fig 4. The zeta potential of the immune complex formation showed an increase of values indicating a high interaction of immune system, mainly for GA condition, Fig 5. The cortisol calibration curve was improved by GA intercalation and the detections were enhanced in concentrated antibody condition, Fig 6. Figure 1

Enzymatic degradation of bacterial cellulose derived carbon nanofibers (BC-CNF) by myeloperoxidase (MPO): Performance evaluation for biosensing

Myeloperoxidase (MPO) is a promising biomarker for early warning of incidence of cardiovascular diseases (CVDs). Among various sensing mechanisms for detection and quantification of MPO, enzymatic degradation of carbon nanoparticles (CNPs) in presence of MPO has unique advantages such as visual change in optical contrast of sample without requiring additional probe chemicals or reagents. While the degradation of various forms of CNPs (including carbon nanotubes and graphene oxide) due to MPO has been reported in literature, the degradation is typically very slow, resulting in response times on the order of multiple hours, and thus presents a significant limitation for CVD diagnosis. Besides, the extent of degradation is highly sensitive to substrate uniformity, necessitating complicated synthesis processes to obtain pristine CNPs. In this work, we have characterized the enzymatic degradation of bacterial cellulose derived carbon nanofibers (BC-CNF) due to MPO in vitro and explored the feasibility of realizing a BC-CNF colorimetric biosensor for measuring MPO activity. The degradation of BC-CNFs is thoroughly characterized using SEM, TEM, Raman and UV–Vis spectroscopy. We report that carboxylic group functionalized BC-CNFs demonstrate noticeable change in optical contrast due to degradation with clinically relevant concentrations of MPO in approximately 1h, and hold great promise for realizing low-cost biosensors for MPO.

Characterizing a New Fluorescent Protein for a Low Limit of Detection Sensing in the Cell-Free System.

Cell-free protein synthesis-based biosensors have been developed as highly accurate, low-cost biosensors. However, since most biomarkers exist at low concentrations in various types of biopsies, the biosensor’s dynamic range must be increased in the system to achieve low limits of detection necessary while deciphering from higher background signals. Many attempts to increase the dynamic range have relied on amplifying the input signal from the analyte, which can lead to complications of false positives. In this study, we aimed to increase the protein synthesis capability of the cell-free protein synthesis system and the output signal of the reporter protein to achieve a lower limit of detection. We utilized a new fluorescent protein, mNeonGreen, which produces a higher output than those commonly used in cell-free biosensors. Optimizations of DNA sequence and the subsequent cell-free protein synthesis reaction conditions allowed characterizing protein expression variability by given DNA template types, reaction environment, and storage additives that cause the greatest time constraint on designing the cell-free biosensor. Finally, we characterized the fluorescence kinetics of mNeonGreen compared to the commonly used reporter protein, superfolder green fluorescent protein. We expect that this finely tuned cell-free protein synthesis platform with the new reporter protein can be used with sophisticated synthetic gene circuitry networks to increase the dynamic range of a cell-free biosensor to reach lower detection limits and reduce the false-positive proportion.

Designed Mini Protein 20 Mimicking Uricase Encapsulated in ZIF-8 as Nanozyme Biosensor for Uric Acid Detection.

This work presents the use of encapsulated mini protein 20 mimicking uricase (mp20)-zeolitic imidazolate framework-8 (ZIF-8) as a bioreceptor for the development of a nanozyme-based electrochemical biosensor for uric acid detection. The electrochemical performance of the biofunctionalized mp20@ZIF-8 on the reduced graphene oxide/screen-printed carbon electrode (rGO/SPCE) was investigated by optimizing operating parameters such as pH, deposition potential, and deposition time using a central composite design-response surface methodology (CCD-RSM). The quadratic regression model was developed to correlate the combination of each variable to the oxidation current density as a response. A significant effect on current response was observed under optimized conditions of pH of 7.4 at −0.35 V deposition potential and 56.56 s deposition time, with p < 0.05 for each interacted factor. The obtained coefficient of determination (R2) value of 0.9992 indicated good agreement with the experimental finding. The developed nanozyme biosensor (mp20@ZIF-8/rGO/SPCE) exhibited high selectivity in the presence of the same fold concentration of interfering species with a detection limit of 0.27 μM, over a concentration range of 1 to 34 μM. The practicality of the tailored biosensor in monitoring uric acid in human serum and urine samples was validated with high-performance liquid chromatography (HPLC) and a commercial uric acid meter. Hence, nanozyme-based is a promising platform that offers a rapid, sensitive, selective, and low-cost biosensor for the non-enzymatic detection of uric acid in biological samples.

Highly sensitive and selective detection of dopamine with boron and sulfur co-doped graphene quantum dots

In this work, we report, the synthesis of Boron and Sulfur co-doped graphene quantum dots (BS-GQDs) and its applicability as a label-free fluorescence sensing probe for the highly sensitive and selective detection of dopamine (DA). Upon addition of DA, the fluorescence intensity of BS-GQDs were effectively quenched over a wide concentration range of DA (0–340 μM) with an ultra-low detection limit of 3.6 μM. The quenching mechanism involved photoinduced electron transfer process from BS-GQDs to dopamine-quinone, produced by the oxidization of DA under alkaline conditions. The proposed sensing mechanism was probed using a detailed study of UV–Vis absorbance, steady state and time resolved fluorescence spectroscopy. The high selectivity of the fluorescent sensor towards DA is established. Our study opens up the possibility of designing a low-cost biosensor which will be suitable for detecting DA in real samples.

Photoelectrochemical nanosensors: An emerging technique for tumor liquid biopsy

As a noninvasive substitute for traditional tissue biopsy, liquid biopsy has emerged as a novel method for the early diagnosis of malignant tumors. In the past decade, low-cost biosensors have been widely studied, and compared to other methods, they have exhibited superior performance in bioanalysis, including liquid biopsies. With the advanced generation of optical and electrochemical biosensors, photoelectrochemical (PEC) biosensors coupled with nanosciences (referred to as PEC nanosensors) have attracted much attention in recent years as they have reached an even higher sensitivity. In this review, the working principle, detection strategy and photoactive nanomaterials of PEC nanosensors were summarized, recent applications of PEC nanosensors for diagnosing different tumor biomolecules in liquid samples were reviewed, and the design processes for each type of PEC nanosensors were especially considered. Finally, some perspectives on developing PEC nanosensors were provided. This review provides systematic retrospective analysis and insights for developing PEC nanosensors in cancer liquid biopsy.

Arduino Based System Design for Measuring Heart Rate and Body Temperature

The aim of this study is to develop a multi-sensor and low-cost biosensor system using Arduino Uno microcontroller board. With this system, heart rate and body temperature can be measured digitally. In this way, the vital values of those with health problems will be monitored regularly, and sudden changes in vital values will be detected immediately. While developing the system, Arduino Uno control card, heart rate sensor and temperature sensor were used. In addition, a software sensitive to detecting changes in heart rate and body temperature has been developed. The system created is open to future additions and improvements. The data obtained through the sensors on the system can be easily tracked by transferring them to the computer over wi-fi. At the same time, it can be improved by placing more sensors in the system according to the purpose of use. As a result of the tests, it has been seen that the system successfully transmits the information received from the sensors to the users in the desired time and gives an audible warning at the determined critical values. Thanks to this system, vital changes in heart rate and body temperature can be accessed remotely and instantly, and it is thought that this will contribute to the field of medicine.

Smartphone-assisted colorimetric biosensor for on-site detection of Cr3+ ion analysis

A simple and low-cost biosensor has been established for the detection of Cr3+ ion by using the image processing function of the smartphone. In the presence of Cr3+ ion, its dependent DNAzyme can specifically cleave the substrate strand, resulting in the release of ligation probe, which will trigger hyperbranched rolling circle amplification (HRCA) to produce hydrogen ions, causing the color change of the solution containing cresol red. A free APP named "Pixel Picker" has been further used to measure the RGB values. It can be well found that the R/G values correlate with Cr3+ ion concentrations with a linear detection range of 0.5 μM–500 μM and a lowest detection limit of 0.12 μM. In addition, the proposed biosensor has been successfully applied to determine Cr3+ ions in various milks. In a word, the constructed smartphone-assisted colorimetric biosensor has the advantages of simplicity, low cost, high specificity and anti-interference ability.

A visual on-site biosensor for low-cost detection of chloramphenicol based on aptamer and split DNAzyme.

Chloramphenicol (CAP) is a kind of broad-spectrum antibiotic, which has been forbidden in food by most countries because of its side effects. In this study, a simple and low-cost biosensor for CAP detection in food was developed. The biosensor consisted of an aptamer specific to CAP and a pair of split probes that could self-assemble as DNAzyme. The detection result could be identified by the naked eye and the visual limit was 10nM CAP. The absorbance of final reaction products at 417nm had a linear relationship with the logarithm of the CAP concentration in a range from 10 to 200nM, and the limit of detection was 87.3pM. The visual analysis by imageJ also showed a linear detection range between 25 and 200nM. The entire detection procedure could be completed in about 1.5h at a cost of about 0.16 dollars per reaction. We believe that the biosensor shows great potential in the rapid and sensitive detection of CAP in food.

Point-of-care detection of pathogenic bacteria based on pregnancy test strips and metal–organic frameworks

Presently, pregnancy test strip (PTS) is one of the most widely used and advanced biomedical testing products. In this study, a portable, sensitive, and low-cost biosensor based on PTS was developed for detecting pathogenic bacteria. In the present study, PTS combined with metal–organic frameworks (MOFs) and hybridization chain reaction (HCR) was used for visual point-of-care (POC) detection of Escherichia coli O157:H7. A single-stranded capture probe (CP) was immobilized onto the surface of magnetic beads (MBs). The CP and an E. coli O157:H7-specific aptamer (AP) formed a double-stranded structure on the surface of MBs. The AP preferentially bound to E. coli O157:H7, thereby exposing some CP and causing their release. Subsequently, the single-stranded CP hybridized with biotin-labeled double-stranded DNA polymers by HCR through the 5ʹ-end sequence of the reporter probe (RP). Numerous MOFs encapsulated with human chorionic gonadotropin (hCG) and modified streptavidin (SA) were then introduced and could be directly and visually detected by PTSs. Under optimal conditions, the method can detect E. coli O157:H7 with a detection limit of 530 CFU/mL. Thus, this method has a high potential for cost-effective, portable, and rapid determination of E. coli O157:H7, especially in resource-limited environments.

Super-Nernstian ion sensitive field-effect transistor exploiting charge screening in WSe2/MoS2 heterostructure

Ion-sensitive field-effect transistors (ISFETs) have gained a lot of attention in recent times as compact, low-cost biosensors with fast response time and label-free detection. Dual gate ISFETs have been shown to enhance detection sensitivity beyond the Nernst limit of 59 mV pH−1 when the back gate dielectric is much thicker than the top dielectric. However, the thicker back-dielectric limits its application for ultrascaled point-of-care devices. In this work, we introduce and demonstrate a pH sensor, with WSe2(top)/MoS2(bottom) heterostructure based double gated ISFET. The proposed device is capable of surpassing the Nernst detection limit and uses thin high-k hafnium oxide as the gate oxide. The 2D atomic layered structure, combined with nanometer-thick top and bottom oxides, offers excellent scalability and linear response with a maximum sensitivity of 362 mV pH−1. We have also used technology computer-aided (TCAD) simulations to elucidate the underlying physics, namely back gate electric field screening through channel and interface charges due to the heterointerface. The proposed mechanism is independent of the dielectric thickness that makes miniaturization of these devices easier. We also demonstrate super-Nernstian behavior with the flipped MoS2(top)/WSe2(bottom) heterostructure ISFET. The results open up a new pathway of 2D heterostructure engineering as an excellent option for enhancing ISFET sensitivity beyond the Nernst limit, for the next-generation of label-free biosensors for single-molecular detection and point-of-care diagnostics.