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A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Nano- and Micro-Technologies in Biosensors".

Deadline for manuscript submissions: closed (31 May 2022) | Viewed by 34555

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Special Issue Editors

Dr. João Henrique Picado Madalena Santos

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Guest Editor

School of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
Interests: downstream processing; alternative solvents; biopharmaceutical process development; nanobiotechnology
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Dibyendu Mondal

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Guest Editor

Centre for Nano and Materials Science, Jain University, Bengaluru 562112, India
Interests: biomass conversion; DNA & protein packaging; biocatalysis; green solvent; water purification

Special Issue Information

Dear Colleagues,

We are pleased to announce an upcoming Special Issue in Biosensors titled “Microbial Toxins and Pathogen Biodetection”. This Special Issue seeks to bring together novel methods that can be useful in the detection of microbial toxins and pathogens for the development of point of care (PoC) devices, novel biosensors, and, in particular, microfluidics devices and nanobiosensors.

Authors are invited to contribute with original research papers, review articles, and short communications that focus on the development and utilization of novel methods to ensure the rapid, simple, and low-cost detection of microbial toxins and pathogens in food, breeding farms, water, the environment, and for clinical applications.

Dr. João H. P. M. Santos
Prof. Dr. Dibyendu Mondal
Guest Editors

Manuscript Submission Information

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. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short 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 thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biosensors 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 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

biodetection
real-time detection
microbial toxins
nanomaterials
biosensors
nanosensors
microorganisms

Published Papers (10 papers)

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Research

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13 pages, 3817 KiB

Open AccessArticle

Evanescent Wave Optical-Fiber Aptasensor for Rapid Detection of Zearalenone in Corn with Unprecedented Sensitivity

by Haixu Zhao, Shang Ren, Zhenzhe Wei and Xinhui Lou

Biosensors 2022, 12(7), 438; https://0-doi-org.brum.beds.ac.uk/10.3390/bios12070438 - 22 Jun 2022

Cited by 5 | Viewed by 2015

Abstract

Zearalenone (ZEN) is a common mycotoxin pollutant found in agricultural products. Aptamers are attractive recognition biomolecules for the development of mycotoxin biosensors. Even though numerous aptasensors have been reported for the detection of ZEN in recent years, many of them suffer from problems including low sensitivity, low specificity, tedious experimental steps, high-cost, and difficulty of automation. We report here the first evanescent wave optical-fiber aptasensor for the detection of ZEN with unprecedented sensitivity, high specificity, low cost, and easy of automation. In our aptasensor, a 40-nt ZEN-specific aptamer (8Z₃₁) is covalently immobilized on the fiber. The 17-nt fluorophore Cy5.5-labeled complementary DNA strand and ZEN competitively bind with the aptamer immobilized on the fiber, enabling the signal-off fluorescent detection of ZEN. The coating of Tween 80 enhanced both the sensitivity and the reproducibility of the aptasensor. The sensor was able to detect ZEN spiked-in the corn flour extract with a semilog linear detection range of 10 pM-10 nM and a limit of detection (LOD, S/N = 3) of 18.4 ± 4.0 pM (equivalent to 29.3 ± 6.4 ng/kg). The LOD is more than 1000-fold lower than the maximum ZEN residue limits set by China (60 μg/kg) and EU (20 μg/kg). The sensor also has extremely high specificity and showed negligible cross-reactivity to other common mycotoxins. In addition, the sensor was able to be regenerated for 28 times, further decreasing its cost. Our sensor holds great potential for practical applications according to its multiple compelling features. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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11 pages, 3017 KiB

Open AccessArticle

A Proof-of-Concept Electrochemical Cytosensor Based on Chlamydomonas reinhardtii Functionalized Carbon Black Screen-Printed Electrodes: Detection of Escherichia coli in Wastewater as a Case Study

by Amina Antonacci, Fabiana Arduini, Raouia Attaallah, Aziz Amine, Maria Teresa Giardi and Viviana Scognamiglio

Biosensors 2022, 12(6), 401; https://0-doi-org.brum.beds.ac.uk/10.3390/bios12060401 - 10 Jun 2022

Cited by 4 | Viewed by 2049

Abstract

Herein, we report a proof-of-concept algal cytosensor for the electrochemical quantification of bacteria in wastewater, exploiting the green photosynthetic alga Chlamydomonas reinhardtii immobilized on carbon black (CB) nanomodified screen-printed electrodes. The CB nanoparticles are used as nanomodifiers, as they are able to sense the oxygen produced by the algae and thus the current increases when algae are exposed to increasing concentrations of bacteria. The sensor was tested on both standard solutions and real wastewater samples for the detection Escherichia coli in a linear range of response from 100 to 2000 CFU/100 mL, showing a limit of detection of 92 CFU/100 mL, in agreement with the maximum E. coli concentration established by the Italian law for wastewater (less than 5000 CFU/100 mL). This bacterium was exploited as a case study target of the algal cytosensor to demonstrate its ability as an early warning analytical system to signal heavy loads of pathogens in waters leaving the wastewater treatment plants. Indeed, the cytosensor is not selective towards E. coli but it is capable of sensing all the bacteria that induce the algae oxygen evolution by exploiting the effect of their interaction. Other known toxicants, commonly present in wastewater, were also analyzed to test the cytosensor selectivity, with any significant effect, apart from atrazine, which is a specific target of the D1 protein of the Chlamydomonas photosystem II. However, the latter can also be detected by chlorophyll fluorescence simultaneously to the amperometric measurements. The matrix effect was evaluated, and the recovery values were calculated as 105 ± 8, 83 ± 7, and 88 ± 7% for 1000 CFU/100 mL of E. coli in Lignano, San Giorgio, and Pescara wastewater samples, respectively. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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16 pages, 3179 KiB

Open AccessArticle

Investigation of Conditions for Capture of Live Legionella pneumophila with Polyclonal and Recombinant Antibodies

by Lucas Paladines, Walid M. Hassen, Juliana Chawich, Stefan Dübel, Simon Lévesque, Jan J. Dubowski and Eric H. Frost

Biosensors 2022, 12(6), 380; https://0-doi-org.brum.beds.ac.uk/10.3390/bios12060380 - 31 May 2022

Cited by 1 | Viewed by 1927

Abstract

Since Legionella pneumophila has caused punctual epidemics through various water systems, the need for a biosensor for fast and accurate detection of pathogenic bacteria in industrial and environmental water has increased. In this report, we evaluated conditions for the capture of live L. pneumophila on a surface by polyclonal antibodies (pAb) and recombinant antibodies (recAb) targeting the bacterial lipopolysaccharide. Using immunoassay and PCR quantification, we demonstrated that, when exposed to live L. pneumophila in PBS or in a mixture containing other non-target bacteria, recAb captured one third fewer L. pneumophila than pAb, but with a 40% lower standard deviation, even when using the same batch of pAb. The presence of other bacteria did not interfere with capture nor increase background by either Ab. Increased reproducibility, as manifested by low standard deviation, is a characteristic that is coveted for biosensing. Hence, the recAb provided a better choice for immune adhesion in biosensors even though it was slightly less sensitive than pAb. Polyclonal or recombinant antibodies can specifically capture large targets such as whole bacteria, and this opens the door to multiple biosensor approaches where any of the components of the bacteria can then be measured for detection or characterisation. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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13 pages, 2285 KiB

Open AccessArticle

Lysine-PEGylated Cytochrome C with Enhanced Shelf-Life Stability

by João H. P. M. Santos, Valker A. Feitosa, Giovanna P. Meneguetti, Gustavo Carretero, João A. P. Coutinho, Sónia P. M. Ventura and Carlota O. Rangel-Yagui

Biosensors 2022, 12(2), 94; https://0-doi-org.brum.beds.ac.uk/10.3390/bios12020094 - 04 Feb 2022

Cited by 5 | Viewed by 2363

Abstract

Cytochrome c (Cyt-c), a small mitochondrial electron transport heme protein, has been employed in bioelectrochemical and therapeutic applications. However, its potential as both a biosensor and anticancer drug is significantly impaired due to poor long-term and thermal stability. To overcome these drawbacks, we developed a site-specific PEGylation protocol for Cyt-c. The PEG derivative used was a 5 kDa mPEG-NHS, and a site-directed PEGylation at the lysine amino-acids was performed. The effects of the pH of the reaction media, molar ratio (Cyt-c:mPEG-NHS) and reaction time were evaluated. The best conditions were defined as pH 7, 1:25 Cyt-c:mPEG-NHS and 15 min reaction time, resulting in PEGylation yield of 45% for Cyt-c-PEG-4 and 34% for Cyt-c-PEG-8 (PEGylated cytochrome c with 4 and 8 PEG molecules, respectively). Circular dichroism spectra demonstrated that PEGylation did not cause significant changes to the secondary and tertiary structures of the Cyt-c. The long-term stability of native and PEGylated Cyt-c forms was also investigated in terms of peroxidative activity. The results demonstrated that both Cyt-c-PEG-4 and Cyt-c-PEG-8 were more stable, presenting higher half-life than unPEGylated protein. In particular, Cyt-c-PEG-8 presented great potential for biomedical applications, since it retained 30–40% more residual activity than Cyt-c over 60-days of storage, at both studied temperatures of 4 °C and 25 °C. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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15 pages, 2884 KiB

Open AccessArticle

Immuno-Dipstick for Colletotrichum gloeosporioides Detection: Towards On-Farm Application

by Fifame Auriane Oussou-Azo, Taiki Futagami and Mun’delanji Catherine M. Vestergaard

Biosensors 2022, 12(2), 49; https://0-doi-org.brum.beds.ac.uk/10.3390/bios12020049 - 18 Jan 2022

Cited by 1 | Viewed by 2855

Abstract

Early and quick detection of pathogens are crucial for managing the spread of infections in the biomedical, biosafety, food, and agricultural fields. While molecular diagnostics can offer the specificity and reliability in acute infectious diseases, detection of pathogens is often slowed down by the current benchtop molecular diagnoses, which are time consuming, labor intensive, and lack the mobility for application at the point-of-need. In this work, we developed a complete on-farm use detection protocol for the plant-devastating anthracnose agent: Colletotrichum gloeosporioides. Our methods combined a simplified DNA extraction on paper that is compatible with loop-mediated isothermal amplification (LAMP), coupled with paper-based immunoassay lateral flow sensing. Our results offer simple, quick, easy, and a minimally instrumented toolkit for Colletotrichum gloeosporioides detection. This scalable and adaptable platform is a valuable alternative to traditional sensing systems towards on-the-go pathogen detection in food and agriculture, biomedical, and other fields. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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24 pages, 5994 KiB

Open AccessEditor’s ChoiceArticle

Bacteriophage-Based Biosensing of Pseudomonas aeruginosa: An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains

by Liliam K. Harada, Waldemar Bonventi Júnior, Erica C. Silva, Thais J. Oliveira, Fernanda C. Moreli, José M. Oliveira Júnior, Matthieu Tubino, Marta M. D. C. Vila and Victor M. Balcão

Biosensors 2021, 11(4), 124; https://0-doi-org.brum.beds.ac.uk/10.3390/bios11040124 - 15 Apr 2021

Cited by 10 | Viewed by 3286

Abstract

During the last decennium, it has become widely accepted that ubiquitous bacterial viruses, or bacteriophages, exert enormous influences on our planet’s biosphere, killing between 4–50% of the daily produced bacteria and constituting the largest genetic diversity pool on our planet. Currently, bacterial infections linked to healthcare services are widespread, which, when associated with the increasing surge of antibiotic-resistant microorganisms, play a major role in patient morbidity and mortality. In this scenario, Pseudomonas aeruginosa alone is responsible for ca. 13–15% of all hospital-acquired infections. The pathogen P. aeruginosa is an opportunistic one, being endowed with metabolic versatility and high (both intrinsic and acquired) resistance to antibiotics. Bacteriophages (or phages) have been recognized as a tool with high potential for the detection of bacterial infections since these metabolically inert entities specifically attach to, and lyse, bacterial host cells, thus, allowing confirmation of the presence of viable cells. In the research effort described herein, three different phages with broad lytic spectrum capable of infecting P. aeruginosa were isolated from environmental sources. The isolated phages were elected on the basis of their ability to form clear and distinctive plaques, which is a hallmark characteristic of virulent phages. Next, their structural and functional stabilization was achieved via entrapment within the matrix of porous alginate, biopolymeric, and bio-reactive, chromogenic hydrogels aiming at their use as sensitive matrices producing both color changes and/or light emissions evolving from a reaction with (released) cytoplasmic moieties, as a bio-detection kit for P. aeruginosa cells. Full physicochemical and biological characterization of the isolated bacteriophages was the subject of a previous research paper. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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Review

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15 pages, 1028 KiB

Open AccessEditor’s ChoiceReview

Transcription Factor-Based Biosensors for Detecting Pathogens

by Yangwon Jeon, Yejin Lee, Keugtae Kim, Geupil Jang and Youngdae Yoon

Biosensors 2022, 12(7), 470; https://0-doi-org.brum.beds.ac.uk/10.3390/bios12070470 - 29 Jun 2022

Cited by 5 | Viewed by 2917

Abstract

Microorganisms are omnipresent and inseparable from our life. Many of them are beneficial to humans, while some are not. Importantly, foods and beverages are susceptible to microbial contamination, with their toxins causing illnesses and even death in some cases. Therefore, monitoring and detecting harmful microorganisms are critical to ensuring human health and safety. For several decades, many methods have been developed to detect and monitor microorganisms and their toxicants. Conventionally, nucleic acid analysis and antibody-based analysis were used to detect pathogens. Additionally, diverse chromatographic methods were employed to detect toxins based on their chemical and structural properties. However, conventional techniques have several disadvantages concerning analysis time, sensitivity, and expense. With the advances in biotechnology, new approaches to detect pathogens and toxins have been reported to compensate for the disadvantages of conventional analysis from different research fields, including electrochemistry, nanotechnology, and molecular biology. Among them, we focused on the recent studies of transcription factor (TF)-based biosensors to detect microorganisms and discuss their perspectives and applications. Additionally, the other biosensors for detecting microorganisms reported in recent studies were also introduced in this review. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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36 pages, 2089 KiB

Open AccessReview

Point-of-Care Diagnostics for Farm Animal Diseases: From Biosensors to Integrated Lab-on-Chip Devices

by Georgios Manessis, Athanasios I. Gelasakis and Ioannis Bossis

Biosensors 2022, 12(7), 455; https://0-doi-org.brum.beds.ac.uk/10.3390/bios12070455 - 26 Jun 2022

Cited by 15 | Viewed by 4248

Abstract

Zoonoses and animal diseases threaten human health and livestock biosecurity and productivity. Currently, laboratory confirmation of animal disease outbreaks requires centralized laboratories and trained personnel; it is expensive and time-consuming, and it often does not coincide with the onset or progress of diseases. Point-of-care (POC) diagnostics are rapid, simple, and cost-effective devices and tests, that can be directly applied on field for the detection of animal pathogens. The development of POC diagnostics for use in human medicine has displayed remarkable progress. Nevertheless, animal POC testing has not yet unfolded its full potential. POC devices and tests for animal diseases face many challenges, such as insufficient validation, simplicity, and portability. Emerging technologies and advanced materials are expected to overcome some of these challenges and could popularize animal POC testing. This review aims to: (i) present the main concepts and formats of POC devices and tests, such as lateral flow assays and lab-on-chip devices; (ii) summarize the mode of operation and recent advances in biosensor and POC devices for the detection of farm animal diseases; (iii) present some of the regulatory aspects of POC commercialization in the EU, USA, and Japan; and (iv) summarize the challenges and future perspectives of animal POC testing. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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22 pages, 2542 KiB

Open AccessReview

Current Challenges and Future Trends of Enzymatic Paper-Based Point-of-Care Testing for Diabetes Mellitus Type 2

by Margarita Ortiz-Martínez, Raquel Flores-DelaToba, Mirna González-González and Marco Rito-Palomares

Biosensors 2021, 11(12), 482; https://0-doi-org.brum.beds.ac.uk/10.3390/bios11120482 - 27 Nov 2021

Cited by 8 | Viewed by 4099

Abstract

A point-of-care (POC) can be defined as an in vitro diagnostic test that can provide results within minutes. It has gained enormous attention as a promising tool for biomarkers detection and diagnosis, as well as for screening of chronic noncommunicable diseases such as diabetes mellitus. Diabetes mellitus type 2 is one of the metabolic disorders that has grown exponentially in recent years, becoming one of the greatest challenges to health systems. Early detection and accurate diagnosis of this disorder are essential to provide adequate treatments. However, efforts to reduce incidence should remain not only in these stages but in developing continuous monitoring strategies. Diabetes-monitoring tools must be accessible and affordable; thus, POC platforms are attractive, especially paper-based ones. Paper-based POCs are simple and portable, can use different matrixes, do not require highly trained staff, and are less expensive than other platforms. These advantages enhance the viability of its application in low-income countries and hard-to-reach zones. This review aims to present a critical summary of the main components required to create a sensitive and affordable enzymatic paper-based POC, as well as an oriented analysis to highlight the main limitations and challenges of current POC devices for diabetes type 2 monitoring and future research opportunities in the field. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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33 pages, 9111 KiB

Open AccessReview

Biosensors Coupled with Signal Amplification Technology for the Detection of Pathogenic Bacteria: A Review

by Fengchun Huang, Yingchao Zhang, Jianhan Lin and Yuanjie Liu

Biosensors 2021, 11(6), 190; https://0-doi-org.brum.beds.ac.uk/10.3390/bios11060190 - 09 Jun 2021

Cited by 37 | Viewed by 6849

Abstract

Foodborne disease caused by foodborne pathogens is a very important issue in food safety. Therefore, the rapid screening and sensitive detection of foodborne pathogens is of great significance for ensuring food safety. At present, many research works have reported the application of biosensors and signal amplification technologies to achieve the rapid and sensitive detection of pathogenic bacteria. Thus, this review summarized the use of biosensors coupled with signal amplification technology for the detection of pathogenic bacteria, including (1) the development, concept, and principle of biosensors; (2) types of biosensors, such as electrochemical biosensors, optical biosensors, microfluidic biosensors, and so on; and (3) different kinds of signal amplification technologies applied in biosensors, such as enzyme catalysis, nucleic acid chain reaction, biotin-streptavidin, click chemistry, cascade reaction, nanomaterials, and so on. In addition, the challenges and future trends for pathogenic bacteria based on biosensor and signal amplification technology were also discussed and summarized. Full article

(This article belongs to the Special Issue Microbial Toxins and Pathogen Biodetection)

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