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Special Issue "Biosensors for Pathogen Detection"

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A special issue of Sensors (ISSN 1424-8220). This special issue belongs to the section "Biosensors".

Deadline for manuscript submissions: closed (28 February 2015)

Special Issue Editor

Guest Editor
Prof. Dr. Stephane Evoy

Department of Electrical and Computer Engineering, University of Alberta, AB T6G 2V4, Edmonton, Canada
Website | E-Mail
Fax: +1 780 492 1811
Interests: nanomechanical and piezoelectric devices for biosensing applications; bacteriophage-based platforms for detection of bacteria

Special Issue Information

Dear Colleagues,

The increased need of improved approaches to pathogen detection for human health care and environmental monitoring has prompted a sustained growth of novel biosensing technologies. A biosensor consists of a miniaturized analytical device integrating biological probes such as enzymes, antibodies, nucleic acids, peptides, viruses, etc., with a transduction platform able to pick up attachment events between the probe and the target. Transduction platforms include all types of physical phenomena including optical, mechanical, piezoelectric, amperometric, etc. Such biosensors offer interesting features such as real-time, on-site, and multiplexed detection of multiple targets through judicious selection of probes. They are thus poised to enable fast and accurate testing platforms in fields such as clinical diagnostics, bioprocess monitoring, environmental monitoring, agricultural product processing, as well as food and water safety. This Special Issue will focus on recent and novel technologies related to both biological probes and transduction systems.

Prof. Dr. Stephane Evoy
Guest Editor

Submission

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. Papers will be published continuously (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are refereed through a peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Sensors is an international peer-reviewed Open Access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 1800 CHF (Swiss Francs).

Keywords

  • pathogens
  • bacteria
  • viruses
  • biosensors
  • food safety
  • water safety
  • diagnosis
  • antibodies
  • agricultural products
  • infectious diseases

Published Papers (12 papers)

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Research

Jump to: Review, Other

Open AccessArticle Highly Sensitive Bacteria Quantification Using Immunomagnetic Separation and Electrochemical Detection of Guanine-Labeled Secondary Beads
Sensors 2015, 15(5), 12034-12052; doi:10.3390/s150512034
Received: 23 March 2015 / Accepted: 7 May 2015 / Published: 22 May 2015
Cited by 3 | PDF Full-text (9047 KB) | HTML Full-text | XML Full-text
Abstract
In this paper, we report the ultra-sensitive indirect electrochemical detection of E. coli O157:H7 using antibody functionalized primary (magnetic) beads for capture and polyguanine (polyG) oligonucleotide functionalized secondary (polystyrene) beads as an electrochemical tag. Vacuum filtration in combination with E. coli O157:H7 specific
[...] Read more.
In this paper, we report the ultra-sensitive indirect electrochemical detection of E. coli O157:H7 using antibody functionalized primary (magnetic) beads for capture and polyguanine (polyG) oligonucleotide functionalized secondary (polystyrene) beads as an electrochemical tag. Vacuum filtration in combination with E. coli O157:H7 specific antibody modified magnetic beads were used for extraction of E. coli O157:H7 from 100 mL samples. The magnetic bead conjugated E. coli O157:H7 cells were then attached to polyG functionalized secondary beads to form a sandwich complex (magnetic bead/E. coli secondary bead). While the use of magnetic beads for immuno-based capture is well characterized, the use of oligonucleotide functionalized secondary beads helps combine amplification and potential multiplexing into the system. The antibody functionalized secondary beads can be easily modified with a different antibody to detect other pathogens from the same sample and enable potential multiplexing. The polyGs on the secondary beads enable signal amplification up to 10\(^{8}\) guanine tags per secondary bead (\(7.5\times10^{6}\) biotin-FITC per secondary bead, 20 guanines per oligonucleotide) bound to the target (E. coli). A single-stranded DNA probe functionalized reduced graphene oxide modified glassy carbon electrode was used to bind the polyGs on the secondary beads. Fluorescent imaging was performed to confirm the hybridization of the complex to the electrode surface. Differential pulse voltammetry (DPV) was used to quantify the amount of polyG involved in the hybridization event with tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)\(_{3}^{2+}\)) as the mediator. The amount of polyG signal can be correlated to the amount of E. coli O157:H7 in the sample. The method was able to detect concentrations of E. coli O157:H7 down to 3 CFU/100 mL, which is 67 times lower than the most sensitive technique reported in literature. The signal to noise ratio for this work was 3. We also demonstrate the use of the protocol for detection of E. coli O157:H7 seeded in waste water effluent samples. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Open AccessArticle Sensor Node for Remote Monitoring of Waterborne Disease-Causing Bacteria
Sensors 2015, 15(5), 10569-10579; doi:10.3390/s150510569
Received: 24 March 2015 / Revised: 20 April 2015 / Accepted: 27 April 2015 / Published: 5 May 2015
Cited by 3 | PDF Full-text (2450 KB) | HTML Full-text | XML Full-text
Abstract
A sensor node for sampling water and checking for the presence of harmful bacteria such as E. coli in water sources was developed in this research. A chromogenic enzyme substrate assay method was used to easily detect coliform bacteria by monitoring the color
[...] Read more.
A sensor node for sampling water and checking for the presence of harmful bacteria such as E. coli in water sources was developed in this research. A chromogenic enzyme substrate assay method was used to easily detect coliform bacteria by monitoring the color change of the sampled water mixed with a reagent. Live webcam image streaming to the web browser of the end user with a Wi-Fi connected sensor node shows the water color changes in real time. The liquid can be manipulated on the web-based user interface, and also can be observed by webcam feeds. Image streaming and web console servers run on an embedded processor with an expansion board. The UART channel of the expansion board is connected to an external Arduino board and a motor driver to control self-priming water pumps to sample the water, mix the reagent, and remove the water sample after the test is completed. The sensor node can repeat water testing until the test reagent is depleted. The authors anticipate that the use of the sensor node developed in this research can decrease the cost and required labor for testing samples in a factory environment and checking the water quality of local water sources in developing countries. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Open AccessArticle Comparison of Whole-Cell SELEX Methods for the Identification of Staphylococcus Aureus-Specific DNA Aptamers
Sensors 2015, 15(4), 8884-8897; doi:10.3390/s150408884
Received: 6 February 2015 / Revised: 28 March 2015 / Accepted: 9 April 2015 / Published: 15 April 2015
Cited by 6 | PDF Full-text (1204 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Whole-cell Systemic Evolution of Ligands by Exponential enrichment (SELEX) is the process by which aptamers specific to target cells are developed. Aptamers selected by whole-cell SELEX have high affinity and specificity for bacterial surface molecules and live bacterial targets. To identify DNA aptamers
[...] Read more.
Whole-cell Systemic Evolution of Ligands by Exponential enrichment (SELEX) is the process by which aptamers specific to target cells are developed. Aptamers selected by whole-cell SELEX have high affinity and specificity for bacterial surface molecules and live bacterial targets. To identify DNA aptamers specific to Staphylococcus aureus, we applied our rapid whole-cell SELEX method to a single-stranded ssDNA library. To improve the specificity and selectivity of the aptamers, we designed, selected, and developed two categories of aptamers that were selected by two kinds of whole-cell SELEX, by mixing and combining FACS analysis and a counter-SELEX process. Using this approach, we have developed a biosensor system that employs a high affinity aptamer for detection of target bacteria. FAM-labeled aptamer sequences with high binding to S. aureus, as determined by fluorescence spectroscopic analysis, were identified, and aptamer A14, selected by the basic whole-cell SELEX using a once-off FACS analysis, and which had a high binding affinity and specificity, was chosen. The binding assay was evaluated using FACS analysis. Our study demonstrated the development of a set of whole-cell SELEX derived aptamers specific to S. aureus; this approach can be used in the identification of other bacteria. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Open AccessCommunication Rapid Detection of Ebola Virus with a Reagent-Free, Point-of-Care Biosensor
Sensors 2015, 15(4), 8605-8614; doi:10.3390/s150408605
Received: 27 February 2015 / Revised: 30 March 2015 / Accepted: 9 April 2015 / Published: 14 April 2015
Cited by 6 | PDF Full-text (2081 KB) | HTML Full-text | XML Full-text
Abstract
Surface acoustic wave (SAW) sensors can rapidly detect Ebola antigens at the point-of-care without the need for added reagents, sample processing, or specialized personnel. This preliminary study demonstrates SAW biosensor detection of the Ebola virus in a concentration-dependent manner. The detection limit with
[...] Read more.
Surface acoustic wave (SAW) sensors can rapidly detect Ebola antigens at the point-of-care without the need for added reagents, sample processing, or specialized personnel. This preliminary study demonstrates SAW biosensor detection of the Ebola virus in a concentration-dependent manner. The detection limit with this methodology is below the average level of viremia detected on the first day of symptoms by PCR. We observe a log-linear sensor response for highly fragmented Ebola viral particles, with a detection limit corresponding to 1.9 × 104 PFU/mL prior to virus inactivation. We predict greatly improved sensitivity for intact, infectious Ebola virus. This point-of-care methodology has the potential to detect Ebola viremia prior to symptom onset, greatly enabling infection control and rapid treatment. This biosensor platform is powered by disposable AA batteries and can be rapidly adapted to detect other emerging diseases in austere conditions. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Figures

Open AccessArticle A Portable Automatic Endpoint Detection System for Amplicons of Loop Mediated Isothermal Amplification on Microfluidic Compact Disk Platform
Sensors 2015, 15(3), 5376-5389; doi:10.3390/s150305376
Received: 16 December 2014 / Revised: 19 January 2015 / Accepted: 28 January 2015 / Published: 5 March 2015
Cited by 3 | PDF Full-text (2067 KB) | HTML Full-text | XML Full-text
Abstract
In recent years, many improvements have been made in foodborne pathogen detection methods to reduce the impact of food contamination. Several rapid methods have been developed with biosensor devices to improve the way of performing pathogen detection. This paper presents an automated endpoint
[...] Read more.
In recent years, many improvements have been made in foodborne pathogen detection methods to reduce the impact of food contamination. Several rapid methods have been developed with biosensor devices to improve the way of performing pathogen detection. This paper presents an automated endpoint detection system for amplicons generated by loop mediated isothermal amplification (LAMP) on a microfluidic compact disk platform. The developed detection system utilizes a monochromatic ultraviolet (UV) emitter for excitation of fluorescent labeled LAMP amplicons and a color sensor to detect the emitted florescence from target. Then it processes the sensor output and displays the detection results on liquid crystal display (LCD). The sensitivity test has been performed with detection limit up to 2.5 × 10−3 ng/µL with different DNA concentrations of Salmonella bacteria. This system allows a rapid and automatic endpoint detection which could lead to the development of a point-of-care diagnosis device for foodborne pathogens detection in a resource-limited environment. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Figures

Open AccessArticle Rapid and Efficient Estimation of Pea Resistance to the Soil-Borne Pathogen Fusarium oxysporum by Infrared Imaging
Sensors 2015, 15(2), 3988-4000; doi:10.3390/s150203988
Received: 12 November 2014 / Accepted: 30 January 2015 / Published: 9 February 2015
Cited by 1 | PDF Full-text (1145 KB) | HTML Full-text | XML Full-text
Abstract
Fusarium wilts are widespread diseases affecting most agricultural crops. In absence of efficient alternatives, sowing resistant cultivars is the preferred approach to control this disease. However, actual resistance sources are often overcome by new pathogenic races, forcing breeders to continuously search for novel
[...] Read more.
Fusarium wilts are widespread diseases affecting most agricultural crops. In absence of efficient alternatives, sowing resistant cultivars is the preferred approach to control this disease. However, actual resistance sources are often overcome by new pathogenic races, forcing breeders to continuously search for novel resistance sources. Selection of resistant accessions, mainly based on the evaluation of symptoms at timely intervals, is highly time-consuming. Thus, we tested the potential of an infra-red imaging system in plant breeding to speed up this process. For this, we monitored the changes in surface leaf temperature upon infection by F. oxysporum f. sp. pisi in several pea accessions with contrasting response to Fusarium wilt under a controlled environment. Using a portable infra-red imaging system we detected a significant temperature increase of at least 0.5 °C after 10 days post-inoculation in the susceptible accessions, while the resistant accession temperature remained at control level. The increase in leaf temperature at 10 days post-inoculation was positively correlated with the AUDPC calculated over a 30 days period. Thus, this approach allowed the early discrimination between resistant and susceptible accessions. As such, applying infra-red imaging system in breeding for Fusarium wilt resistance would contribute to considerably shorten the process of selection of novel resistant sources. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Open AccessArticle A Novel Ultrasensitive ECL Sensor for DNA Detection Based on Nicking Endonuclease-Assisted Target Recycling Amplification, Rolling Circle Amplification and Hemin/G-Quadruplex
Sensors 2015, 15(2), 2629-2643; doi:10.3390/s150202629
Received: 1 December 2014 / Accepted: 19 January 2015 / Published: 26 January 2015
Cited by 3 | PDF Full-text (2121 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
In this study, we describe a novel universal and highly sensitive strategy for the electrochemiluminescent (ECL) detection of sequence specific DNA at the aM level based on Nt.BbvCI (a nicking endonuclease)-assisted target recycling amplification (TRA), rolling circle amplification (RCA) and hemin/G-quadruplex. The target
[...] Read more.
In this study, we describe a novel universal and highly sensitive strategy for the electrochemiluminescent (ECL) detection of sequence specific DNA at the aM level based on Nt.BbvCI (a nicking endonuclease)-assisted target recycling amplification (TRA), rolling circle amplification (RCA) and hemin/G-quadruplex. The target DNAs can hybridize with self-assembled capture probes and assistant probes to form “Y” junction structures on the electrode surface, thus triggering the execution of a TRA reaction with the aid of Nt.BbvCI. Then, the RCA reaction and the addition of hemin result in the production of numerous hemin/G-quadruplex, which consume the dissolved oxygen in the detection buffer and result in a significant ECL quenching effect toward the O2/S2O82− system. The proposed strategy combines the amplification ability of TRA, RCA and the inherent high sensitivity of the ECL technique, thus enabling low aM (3.8 aM) detection for sequence-specific DNA and a wide linear range from 10.0 aM to 1.0 pM. At the same time, this novel strategy shows high selectivity against single-base mismatch sequences, which makes our novel universal and highly sensitive method a powerful addition to specific DNA sequence detection. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Open AccessArticle Comparison between a Conductometric Biosensor and ELISA in the Evaluation of Johne’s Disease
Sensors 2014, 14(10), 19128-19137; doi:10.3390/s141019128
Received: 11 August 2014 / Revised: 8 October 2014 / Accepted: 9 October 2014 / Published: 15 October 2014
Cited by 1 | PDF Full-text (842 KB) | HTML Full-text | XML Full-text
Abstract
Johne’s disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), is an important gastrointestinal disease of cattle worldwide because of the economic losses encountered in JD-affected herds. These losses include reduction in milk yield in cows, premature culling and reduced carcass weight of
[...] Read more.
Johne’s disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), is an important gastrointestinal disease of cattle worldwide because of the economic losses encountered in JD-affected herds. These losses include reduction in milk yield in cows, premature culling and reduced carcass weight of culled diseased animals. In the U.S. dairy industry, economic losses from reduced productivity associated with JD are estimated to cost between $200 and $250 million annually. The development of non-laboratory-based assays would support more frequent testing of animals for JD and could improve its control. Conductometric biosensors combine immunomigration technology with electronic signal detection and have been adapted for the detection of IgG antibody against MAP. In the present study, a capture membrane with limited variability in the immunomigration channel and an optimal concentration of the secondary anti-bovine antibody used in a previously developed conductometric biosensor were compared with a commercially available antibody detection ELISA in their evaluation of JD, using samples of serum from cattle whose JD status where unknown. There was a moderate strength of agreement (kappa = 0.41) between the two assays. Findings from this preliminary study support the continued development of conductometric biosensors for use in the diagnosis of JD. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)

Review

Jump to: Research, Other

Open AccessReview A Review of Membrane-Based Biosensors for Pathogen Detection
Sensors 2015, 15(6), 14045-14078; doi:10.3390/s150614045
Received: 29 April 2015 / Revised: 29 May 2015 / Accepted: 5 June 2015 / Published: 15 June 2015
Cited by 4 | PDF Full-text (1916 KB) | HTML Full-text | XML Full-text
Abstract
Biosensors are of increasing interest for the detection of bacterial pathogens in many applications such as human, animal and plant health, as well as food and water safety. Membranes and membrane-like structures have been integral part of several pathogen detection platforms. Such structures
[...] Read more.
Biosensors are of increasing interest for the detection of bacterial pathogens in many applications such as human, animal and plant health, as well as food and water safety. Membranes and membrane-like structures have been integral part of several pathogen detection platforms. Such structures may serve as simple mechanical support, function as a part of the transduction mechanism, may be used to filter out or concentrate pathogens, and may be engineered to specifically house active proteins. This review focuses on membrane materials, their associated biosensing applications, chemical linking procedures, and transduction mechanisms. The sensitivity of membrane biosensors is discussed, and the state of the field is evaluated and summarized. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Open AccessReview Modulated Raman Spectroscopy for Enhanced Cancer Diagnosis at the Cellular Level
Sensors 2015, 15(6), 13680-13704; doi:10.3390/s150613680
Received: 14 April 2015 / Revised: 22 May 2015 / Accepted: 24 May 2015 / Published: 11 June 2015
Cited by 5 | PDF Full-text (14079 KB) | HTML Full-text | XML Full-text
Abstract
Raman spectroscopy is emerging as a promising and novel biophotonics tool for non-invasive, real-time diagnosis of tissue and cell abnormalities. However, the presence of a strong fluorescence background is a key issue that can detract from the use of Raman spectroscopy in routine
[...] Read more.
Raman spectroscopy is emerging as a promising and novel biophotonics tool for non-invasive, real-time diagnosis of tissue and cell abnormalities. However, the presence of a strong fluorescence background is a key issue that can detract from the use of Raman spectroscopy in routine clinical care. The review summarizes the state-of-the-art methods to remove the fluorescence background and explores recent achievements to address this issue obtained with modulated Raman spectroscopy. This innovative approach can be used to extract the Raman spectral component from the fluorescence background and improve the quality of the Raman signal. We describe the potential of modulated Raman spectroscopy as a rapid, inexpensive and accurate clinical tool to detect the presence of bladder cancer cells. Finally, in a broader context, we show how this approach can greatly enhance the sensitivity of integrated Raman spectroscopy and microfluidic systems, opening new prospects for portable higher throughput Raman cell sorting. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)
Open AccessReview Development of a HIV-1 Virus Detection System Based on Nanotechnology
Sensors 2015, 15(5), 9915-9927; doi:10.3390/s150509915
Received: 5 March 2015 / Revised: 8 April 2015 / Accepted: 20 April 2015 / Published: 27 April 2015
Cited by 4 | PDF Full-text (2380 KB) | HTML Full-text | XML Full-text
Abstract
Development of a sensitive and selective detection system for pathogenic viral agents is essential for medical healthcare from diagnostics to therapeutics. However, conventional detection systems are time consuming, resource-intensive and tedious to perform. Hence, the demand for sensitive and selective detection system for
[...] Read more.
Development of a sensitive and selective detection system for pathogenic viral agents is essential for medical healthcare from diagnostics to therapeutics. However, conventional detection systems are time consuming, resource-intensive and tedious to perform. Hence, the demand for sensitive and selective detection system for virus are highly increasing. To attain this aim, different aspects and techniques have been applied to develop virus sensor with improved sensitivity and selectivity. Here, among those aspects and techniques, this article reviews HIV virus particle detection systems incorporated with nanotechnology to enhance the sensitivity. This review mainly focused on four different detection system including vertically configured electrical detection based on scanning tunneling microscopy (STM), electrochemical detection based on direct electron transfer in virus, optical detection system based on localized surface plasmon resonance (LSPR) and surface enhanced Raman spectroscopy (SERS) using plasmonic nanoparticle. Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)

Other

Jump to: Research, Review

Open AccessBrief Report Non-Invasive Optical Sensor Based Approaches for Monitoring Virus Culture to Minimize BSL3 Laboratory Entry
Sensors 2015, 15(7), 14864-14870; doi:10.3390/s150714864
Received: 24 March 2015 / Revised: 13 May 2015 / Accepted: 15 June 2015 / Published: 24 June 2015
Cited by 1 | PDF Full-text (1480 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
High titers of infectious viruses for vaccine and diagnostic reference panel development are made by infecting susceptible mammalian cells. Laboratory procedures are strictly performed in a Bio-Safety Level-3 (BSL3) laboratory and each entry and exit involves the use of  disposable Personnel Protective Equipment
[...] Read more.
High titers of infectious viruses for vaccine and diagnostic reference panel development are made by infecting susceptible mammalian cells. Laboratory procedures are strictly performed in a Bio-Safety Level-3 (BSL3) laboratory and each entry and exit involves the use of  disposable Personnel Protective Equipment (PPE) to observe cell culture conditions. Routine PPE use involves significant recurring costs. Alternative non-invasive optical sensor based approaches to remotely monitor cell culture may provide a promising and cost effective approach to monitor infectious virus cultures resulting in lower disruption and costs. We report here the monitoring of high titer cultures of Human Immunodeficiency Virus-1 (HIV-1) and Herpes Simplex Virus-2 (HSV-2) remotely with the use of optical oxygen sensors aseptically placed inside the cell culture vessel. The replacement of culture media for cell and virus propagation and virus load monitoring was effectively performed using this fluorescent sensor and resulted in half the number of visits to the BSL3 lab (five versus ten). Full article
(This article belongs to the Special Issue Biosensors for Pathogen Detection)

Planned Papers

The below list represents only planned manuscripts. Some of these manuscripts have not been received by the Editorial Office yet. Papers submitted to MDPI journals are subject to peer-review.

Type: Review
Title:
A Review of Membrane-Based Biosensors for Pathogen Detection
Authors:
Remko van den Hurk, Stephane Evoy
Affiliations:
Electrical and Computer Engineering, University of Alberta, Canada
Abstract:
Micro- and nanomechanical structures, such as cantilevers, paddle oscillators, beams and membranes, have received sustained attention for a wide range of applications, including sensing, energy harvesting, telecommunications, and information processing. Membranes have specifically been employed in devices such as bio- and chemical, tactile and pressure sensors; actuators; optical devices; plasmonic structures; fuel cells; biomolecules sorters; supercapacitors, lithium ion storage batteries, capacitive micromachined ultrasonic transducers (CMUTs), and acoustic energy harvesters. We, here, specifically review recent advances concerning the realization of functional nanomembranes in applications relevant to pathogen detection. Such membranes have been used as a mechanical support, as part of the transduction mechanism, and on platforms for the sorting of biomolecules and pathogens. These membranes may also be engineered to specifically house active proteins. We will review the different materials and fabrication methods used to create these membrane biosensors, the various transduction and detection methods employed, and outline the future potential in the field.

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