Research

15 pages, 1368 KiB

Open AccessArticle

Flow-Through Electrochemical Biosensor for the Detection of Listeria monocytogenes Using Oligonucleotides

by Cheryl M. Armstrong, Joe Lee, Andrew G. Gehring and Joseph A. Capobianco

Sensors 2021, 21(11), 3754; https://doi.org/10.3390/s21113754 - 28 May 2021

Cited by 7 | Viewed by 2875

Consumption of food contaminated by Listeria monocytogenes can result in Listeriosis, an illness with hospitalization rates of 94% and mortality rates up to 30%. As a result, U.S. regulatory agencies governing food safety retain zero-tolerance policies for L. monocytogenes. However, detection at [...] Read more.

Consumption of food contaminated by Listeria monocytogenes can result in Listeriosis, an illness with hospitalization rates of 94% and mortality rates up to 30%. As a result, U.S. regulatory agencies governing food safety retain zero-tolerance policies for L. monocytogenes. However, detection at such low concentrations often requires strategies such as increasing sample size or culture enrichment. A novel flow-through immunoelectrochemical biosensor has been developed for Escherichia coli O157:H7 detection in 1 L volumes without enrichment. The current work further augments this biosensor’s capabilities to (1) include detection of L. monocytogenes and (2) accommodate genetic detection to help overcome limitations based upon antibody availability and address specificity errors in phenotypic assays. Herein, the conjugation scheme for oligo attachment and the conditions necessary for genetic detection are laid forth while results of the present study demonstrate the sensor’s ability to distinguish L. monocytogenes DNA from L. innocua with a limit of detection of ~2 × 10⁴ cells/mL, which agrees with prior studies. Total time for this assay can be constrained to <2.5 h because a timely culture enrichment period is not necessary. Furthermore, the electrochemical detection assay can be performed with hand-held electronics, allowing this platform to be adopted for near-line monitoring systems. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

18 pages, 2979 KiB

Open AccessArticle

Computational Strategies for the Identification of a Transcriptional Biomarker Panel to Sense Cellular Growth States in Bacillus subtilis

by Yiming Huang, Wendy Smith, Colin Harwood, Anil Wipat and Jaume Bacardit

Sensors 2021, 21(7), 2436; https://doi.org/10.3390/s21072436 - 01 Apr 2021

Cited by 2 | Viewed by 2789

Abstract

A goal of the biotechnology industry is to be able to recognise detrimental cellular states that may lead to suboptimal or anomalous growth in a bacterial population. Our current knowledge of how different environmental treatments modulate gene regulation and bring about physiology adaptations [...] Read more.

A goal of the biotechnology industry is to be able to recognise detrimental cellular states that may lead to suboptimal or anomalous growth in a bacterial population. Our current knowledge of how different environmental treatments modulate gene regulation and bring about physiology adaptations is limited, and hence it is difficult to determine the mechanisms that lead to their effects. Patterns of gene expression, revealed using technologies such as microarrays or RNA-seq, can provide useful biomarkers of different gene regulatory states indicative of a bacterium’s physiological status. It is desirable to have only a few key genes as the biomarkers to reduce the costs of determining the transcriptional state by opening the way for methods such as quantitative RT-PCR and amplicon panels. In this paper, we used unsupervised machine learning to construct a transcriptional landscape model from condition-dependent transcriptome data, from which we have identified 10 clusters of samples with differentiated gene expression profiles and linked to different cellular growth states. Using an iterative feature elimination strategy, we identified a minimal panel of 10 biomarker genes that achieved 100% cross-validation accuracy in predicting the cluster assignment. Moreover, we designed and evaluated a variety of data processing strategies to ensure our methods were able to generate meaningful transcriptional landscape models, capturing relevant biological processes. Overall, the computational strategies introduced in this study facilitate the identification of a detailed set of relevant cellular growth states, and how to sense them using a reduced biomarker panel. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

22 pages, 3188 KiB

Open AccessArticle

Rational Design of a User-Friendly Aptamer/Peptide-Based Device for the Detection of Staphylococcus aureus

by Luca Ronda, Alessandro Tonelli, Elisa Sogne, Ida Autiero, Francesca Spyrakis, Sara Pellegrino, Giorgio Abbiati, Elisa Maffioli, Carsten Schulte, Riccardo Piano, Pietro Cozzini, Andrea Mozzarelli, Stefano Bettati, Francesca Clerici, Paolo Milani, Cristina Lenardi, Gabriella Tedeschi and Maria Luisa Gelmi

Sensors 2020, 20(17), 4977; https://doi.org/10.3390/s20174977 - 02 Sep 2020

Cited by 7 | Viewed by 3491

Abstract

The urgent need to develop a detection system for Staphylococcus aureus, one of the most common causes of infection, is prompting research towards novel approaches and devices, with a particular focus on point-of-care analysis. Biosensors are promising systems to achieve this aim. [...] Read more.

The urgent need to develop a detection system for Staphylococcus aureus, one of the most common causes of infection, is prompting research towards novel approaches and devices, with a particular focus on point-of-care analysis. Biosensors are promising systems to achieve this aim. We coupled the selectivity and affinity of aptamers, short nucleic acids sequences able to recognize specific epitopes on bacterial surface, immobilized at high density on a nanostructured zirconium dioxide surface, with the rational design of specifically interacting fluorescent peptides to assemble an easy-to-use detection device. We show that the displacement of fluorescent peptides upon the competitive binding of S. aureus to immobilized aptamers can be detected and quantified through fluorescence loss. This approach could be also applied to the detection of other bacterial species once aptamers interacting with specific antigens will be identified, allowing the development of a platform for easy detection of a pathogen without requiring access to a healthcare environment. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

12 pages, 2442 KiB

Open AccessArticle

Multiple Bacteria Identification in the Point-of-Care: an Old Method Serving a New Approach

by Sara Viveiros, Mónica Rodrigues, Débora Albuquerque, Sofia A. M. Martins, Susana Cardoso and Verónica C. Martins

Sensors 2020, 20(12), 3351; https://doi.org/10.3390/s20123351 - 12 Jun 2020

Cited by 6 | Viewed by 2624

Abstract

The accurate diagnosis of bacterial infections is of critical importance for effective treatment decisions. Due to the multietiologic nature of most infectious diseases, multiplex assays are essential for diagnostics. However, multiplexability in nucleic acid amplification-based methods commonly resorts to multiple primers and/or multiple [...] Read more.

The accurate diagnosis of bacterial infections is of critical importance for effective treatment decisions. Due to the multietiologic nature of most infectious diseases, multiplex assays are essential for diagnostics. However, multiplexability in nucleic acid amplification-based methods commonly resorts to multiple primers and/or multiple reaction chambers, which increases analysis cost and complexity. Herein, a polymerase chain reaction (PCR) offer method based on a universal pair of primers and an array of specific oligonucleotide probes was developed through the analysis of the bacterial 16S ribosomal RNA gene. The detection system consisted of DNA hybridization over an array of magnetoresistive sensors in a microfabricated biochip coupled to an electronic reader. Immobilized probes interrogated single-stranded biotinylated amplicons and were obtained using asymmetric PCR. Moreover, they were magnetically labelled with streptavidin-coated superparamagnetic nanoparticles. The benchmarking of the system was demonstrated to detect five major bovine mastitis-causing pathogens: Escherichia coli, Klebsiella sp., Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae. All selected probes proved to specifically detect their respective amplicon without significant cross reactivity. A calibration curve was performed for S. agalactiae, which demonstrates demonstrating a limit of detection below 30 fg/µL. Thus, a sensitive and specific multiplex detection assay was established, demonstrating its potential as a bioanalytical device for point-of-care applications. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

11 pages, 3909 KiB

Open AccessArticle

Detection of Escherichia coli O157:H7 Using Automated Immunomagnetic Separation and Enzyme-Based Colorimetric Assay

by Ji Young Park, Kisang Park, Gyeongsik Ok, Hyun-Joo Chang, Tae Jung Park, Sung-Wook Choi and Min-Cheol Lim

Sensors 2020, 20(5), 1395; https://doi.org/10.3390/s20051395 - 04 Mar 2020

Cited by 22 | Viewed by 6132

Abstract

The food industry requires rapid and simple detection methods for preventing harm from pathogenic bacteria. Until now, various technologies used to detect foodborne bacteria were time-consuming and laborious. Therefore, we have developed an automated immunomagnetic separation combined with a colorimetric assay for the [...] Read more.

The food industry requires rapid and simple detection methods for preventing harm from pathogenic bacteria. Until now, various technologies used to detect foodborne bacteria were time-consuming and laborious. Therefore, we have developed an automated immunomagnetic separation combined with a colorimetric assay for the rapid detection of E. coli O157:H7 in food samples. The colorimetric detection method using enzymatic reaction is fascinating because of its simplicity and rapidity and does not need sophisticated devices. Moreover, the proposed procedures for the detection of bacteria in food take less than 3 h including pre-enrichment, separation and detection steps. First, target-specific immunomagnetic beads were introduced to contaminated milk in a pre-enrichment step. Second, the pre-enriched sample solution containing target bacteria bound on immunomagnetic beads was injected into an automated pretreatment system. Subsequently, the immunomagnetic beads along with target bacteria were separated and concentrated into a recovery tube. Finally, released β-galactosidase from E. coli O157:H7 after lysis was reacted with chlorophenol red β-galactopyranoside (CPRG) used as a substrate and the colorimetric change of CPRG was determined by absorbance measuring or the naked eye. By the proposed approach in this study, we could detect 3 × 10² CFU/mL of E. coli O157:H7 from a milk sample within 3 h. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

11 pages, 3372 KiB

Open AccessArticle

Light Energy Conversion Surface with Gold Dendritic Nanoforests/Si Chip for Plasmonic Polymerase Chain Reaction

by Hung Ji Huang, Yu-Cheng Chiang, Chia-Hsien Hsu, Jyh-Jian Chen, Ming-Hua Shiao, Chih-Chieh Yeh, Shu-Ling Huang and Yung-Sheng Lin

Sensors 2020, 20(5), 1293; https://doi.org/10.3390/s20051293 - 27 Feb 2020

Cited by 9 | Viewed by 2621

Abstract

Surfaces with gold dendritic nanoforests (Au DNFs) on Si chips demonstrate broadband-light absorption. This study is the first to utilize localized surface plasmons of Au DNFs/Si chips for polymerase chain reaction (PCR) applications. A convenient halogen lamp was used as the heating source [...] Read more.

Surfaces with gold dendritic nanoforests (Au DNFs) on Si chips demonstrate broadband-light absorption. This study is the first to utilize localized surface plasmons of Au DNFs/Si chips for polymerase chain reaction (PCR) applications. A convenient halogen lamp was used as the heating source to illuminate the Au DNFs/Si chip for PCR. A detection target of Salmonella spp. DNA fragments was reproduced in this plasmonic PCR chip system. By semi-quantitation in gel electrophoresis analysis, the plasmonic PCR with 30 cycles and a largely reduced processing time provided results comparable with those of a commercial PCR thermal cycler with 40 cycles in more than 1 h. In the presence of an Au DNFs/Si chip, the plasmonic PCR provides superior results in a short processing time. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

Review

Jump to: Research, Other

23 pages, 3373 KiB

Open AccessReview

Advanced Methods for Detection of Bacillus cereus and Its Pathogenic Factors

by Nalini Ramarao, Seav-Ly Tran, Marco Marin and Jasmina Vidic

Sensors 2020, 20(9), 2667; https://doi.org/10.3390/s20092667 - 07 May 2020

Cited by 65 | Viewed by 11332

Abstract

Bacillus cereus is an opportunistic foodborne pathogen causing food intoxication and infectious diseases. Different toxins and pathogenic factors are responsible for diarrheal syndrome, like nonhemolytic enterotoxin Nhe, hemolytic enterotoxin Hbl, enterotoxin FM and cytotoxin K, while emetic syndrome is caused by the depsipeptide [...] Read more.

Bacillus cereus is an opportunistic foodborne pathogen causing food intoxication and infectious diseases. Different toxins and pathogenic factors are responsible for diarrheal syndrome, like nonhemolytic enterotoxin Nhe, hemolytic enterotoxin Hbl, enterotoxin FM and cytotoxin K, while emetic syndrome is caused by the depsipeptide cereulide toxin. The traditional method of B. cereus detection is based on the bacterial culturing onto selective agars and cells enumeration. In addition, molecular and chemical methods are proposed for toxin gene profiling, toxin quantification and strain screening for defined virulence factors. Finally, some advanced biosensors such as phage-based, cell-based, immunosensors and DNA biosensors have been elaborated to enable affordable, sensitive, user-friendly and rapid detection of specific B. cereus strains. This review intends to both illustrate the state of the B. cereus diagnostic field and to highlight additional research that is still at the development level. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Graphical abstract

32 pages, 6046 KiB

Open AccessReview

Review of Electrochemical DNA Biosensors for Detecting Food Borne Pathogens

by Qiaoyun Wu, Yunzhe Zhang, Qian Yang, Ning Yuan and Wei Zhang

Sensors 2019, 19(22), 4916; https://doi.org/10.3390/s19224916 - 12 Nov 2019

Cited by 67 | Viewed by 9695

Abstract

The vital importance of rapid and accurate detection of food borne pathogens has driven the development of biosensor to prevent food borne illness outbreaks. Electrochemical DNA biosensors offer such merits as rapid response, high sensitivity, low cost, and ease of use. This review [...] Read more.

The vital importance of rapid and accurate detection of food borne pathogens has driven the development of biosensor to prevent food borne illness outbreaks. Electrochemical DNA biosensors offer such merits as rapid response, high sensitivity, low cost, and ease of use. This review covers the following three aspects: food borne pathogens and conventional detection methods, the design and fabrication of electrochemical DNA biosensors and several techniques for improving sensitivity of biosensors. We highlight the main bioreceptors and immobilizing methods on sensing interface, electrochemical techniques, electrochemical indicators, nanotechnology, and nucleic acid-based amplification. Finally, in view of the existing shortcomings of electrochemical DNA biosensors in the field of food borne pathogen detection, we also predict and prospect future research focuses from the following five aspects: specific bioreceptors (improving specificity), nanomaterials (enhancing sensitivity), microfluidic chip technology (realizing automate operation), paper-based biosensors (reducing detection cost), and smartphones or other mobile devices (simplifying signal reading devices). Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

Other

Jump to: Research, Review

11 pages, 8097 KiB

Open AccessLetter

Integrated, Automated, Fast PCR System for Point-Of-Care Molecular Diagnosis of Bacterial Infection

by Dongkyu Lee, Deawook Kim, Jounghyuk Han, Jongsu Yun, Kang-Ho Lee, Gyu Man Kim, Ohwon Kwon and Jaejong Lee

Sensors 2021, 21(2), 377; https://doi.org/10.3390/s21020377 - 07 Jan 2021

Cited by 8 | Viewed by 4113

Abstract

We developed an integrated PCR system that performs automated sample preparation and fast polymerase chain reaction (PCR) for application in point-of care (POC) testing. This system is assembled from inexpensive 3D-printing parts, off-the-shelf electronics and motors. Molecular detection requires a series of procedures [...] Read more.

We developed an integrated PCR system that performs automated sample preparation and fast polymerase chain reaction (PCR) for application in point-of care (POC) testing. This system is assembled from inexpensive 3D-printing parts, off-the-shelf electronics and motors. Molecular detection requires a series of procedures including sample preparation, amplification, and fluorescence intensity analysis. The system can perform automated DNA sample preparation (extraction, separation and purification) in ≤5 min. The variance of the automated sample preparation was clearly lower than that achieved using manual DNA extraction. Fast thermal ramp cycles were generated by a customized thermocycler designed to automatically transport samples between heating and cooling blocks. Despite the large sample volume (50 μL), rapid two-step PCR amplification completed 40 cycles in ≤13.8 min. Variations in fluorescence intensity were measured by analyzing fluorescence images. As proof of concept of this system, we demonstrated the rapid DNA detection of pathogenic bacteria. We also compared the sensitivity of this system with that of a commercial device during the automated extraction and fast PCR of Salmonella bacteria. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

13 pages, 3613 KiB

Open AccessLetter

Performances and Biosensing Mechanisms of Interdigitated Capacitive Sensors Based on the Hetero-mixture of SnO₂ and In₂O₃

by Akhil Chandran Mukkattu Kuniyil, Janez Zavašnik, Željka Cvejić, Sohail Sarang, Mitar Simić, Vladimir V. Srdić and Goran M. Stojanović

Sensors 2020, 20(21), 6323; https://doi.org/10.3390/s20216323 - 06 Nov 2020

Cited by 3 | Viewed by 3017

Abstract

This study aims to discuss the synthesis and fabrication of SnO₂-In₂O₃-based thick-films and their biosensing applications. The structural characterization of SnO₂-In₂O₃ nanocomposites was performed using X-ray diffraction, Raman spectroscopy and transmission electron [...] Read more.

This study aims to discuss the synthesis and fabrication of SnO₂-In₂O₃-based thick-films and their biosensing applications. The structural characterization of SnO₂-In₂O₃ nanocomposites was performed using X-ray diffraction, Raman spectroscopy and transmission electron microscopy. Furthermore, the screen-printing technology was used in the fabrication of conductive electrodes to form an interdigitated capacitive structure, and the sensor layer based on the mixture of SnO₂ and In₂O₃. Moreover, the sensing performance of the developed structure was tested using Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) bacteria. In addition, the validation of sensing characteristics was performed by electrochemical impedance spectroscopic and self-resonant frequency analysis. Finally, the sensing properties were analyzed for two consecutive days, and changes in both P. aeruginosa and S. aureus pathogens growing media were also studied. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

11 pages, 3465 KiB

Open AccessLetter

Interdigitated and Wave-Shaped Electrode-Based Capacitance Sensor for Monitoring Antibiotic Effects

by Jinsoo Park, Yonghyun Lee, Youjin Hwang and Sungbo Cho

Sensors 2020, 20(18), 5237; https://doi.org/10.3390/s20185237 - 14 Sep 2020

Cited by 4 | Viewed by 2674

Abstract

Label-free and real-time monitoring of the bacterial viability is essential for the accurate and sensitive characterization of the antibiotic effects. In the present study, we investigated the feasibility of the interdigitated and wave-shaped electrode (IWE) for monitoring the effect of tetracycline or kanamycin [...] Read more.

Label-free and real-time monitoring of the bacterial viability is essential for the accurate and sensitive characterization of the antibiotic effects. In the present study, we investigated the feasibility of the interdigitated and wave-shaped electrode (IWE) for monitoring the effect of tetracycline or kanamycin on Staphylococcus aureus (S. aureus) and methicillin-resistant S.aureus (MRSA). The electrical impedance spectra of the IWE immersed in the culture media for bacterial growth were characterized in a frequency range of 10 Hz to 1 kHz. The capacitance index (CI) (capacitance change relevant with the bacterial viability) was used to monitor the antibiotic effects on the S. aureus and MRSA in comparison to the traditional methods (disk diffusion test and optical density (OD) measurement). The experimental results showed that the percentage of change in CI (PCI) for the antibiotic effect on MRSA was increased by 51.58% and 57.83% in kanamycin and control, respectively. In contrast, the PCI value decreased by 0.25% for tetracycline, decreased by 52.63% and 37.66% in the cases of tetracycline and kanamycin-treated S. aureus, and increased 2.79% in the control, respectively. This study demonstrated the feasibility of the IWE-based capacitance sensor for the label-free and real-time monitoring of the antibiotic effects on S. aureus and MRSA. Full article

(This article belongs to the Special Issue Advanced Biosensors for Bacterial Detection)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Advanced Biosensors for Bacterial Detection

Share This Special Issue

Special Issue Editor

Special Issue Information

Keywords

Published Papers (11 papers)

Research

Review

Other

Further Information

Guidelines

MDPI Initiatives

Follow MDPI